FR2670683A1 - METHOD AND SYSTEM, COMPUTER ASSISTED TO ENHANCE ELECTROPHORESIS SEPARATION OF MACROMOLECULES, IN PARTICULAR DNA. - Google Patents

Abstract

A system for improving the electrophoretic separation of macromolecules is provided. It relates to a system (100) which comprises a knowledge base (120) derived from calibration data obtained by separation of macromolecules, in particular DNA, of known size, under various conditions chosen for the optimization of the separations. The user enters the smallest size and the largest size of DNA macromolecules to be separated. Based on this information, the system (100) automatically determines the parameters of the electrophoresis to improve its resolution. Application to the separations of macromolecules by electrophoresis.

Description

The present invention relates generally to gel electrophoresis

  and more specifically it relates to an electrophoresis control system with an electric field

  homogeneous locked to its outline.

  The conventional method used for separation,

  identification and purification of deoxyribonucleic acid

  nucleic acid (hereinafter referred to as DNA) and other molecules is electrophoresis on agarose gels. This technique allows the efficient and rapid resolution of DNA fragment mixtures that can not be conveniently separated by other purification technologies A gel containing a DNA sample is subjected to an electric field This causes the migration

  individual macromolecules of the DNA in the gel.

  As the different DNA fragments migrate at different rates, visible "bands" of DNA form in the gel. The location of the individual DNA fragments in the

  gel can be read directly from the bands.

  Particular characteristics of DNA, particularly

  Its size, charge and conformation influence its rate of migration and thus the separation that can be obtained. For example, linear and double DNA molecules move in gelled materials at rates that are, for example, inversely proportional.

  logarithm at base ten of their molecular mass

  less, DNA macromolecules of identical molecular weights but of different conformations, for example closed circular, shrunken circular and linear, migrate in an agarose gel at speeds

different.

  In addition to the characteristics of DNA,

  electrophoresis meters also affect the migration (mobility) speed of macromolecules DNA fragments of given size migrate at different rates in gels containing different concentrations of agarose The variation in concentration allows the resolution of d a wide range of DNA fragment sizes The intensity of the applied field (voltage gradient), the ionic concentration of the buffer solution, and the density of superhelical bends in the closed circular form of DNA also have an influence. on migration At low voltages, the mobility (cm / h) of linear DNA fragments is proportional to the applied voltage. However, this linear relationship no longer exists at the highest potential for high molecular weight fragments. effective separation range for determined agarose gels decreases

when the voltage increases.

  Better separation of the macro-

  by optimizing certain parameters. For example, pulsed-field gel electrophoresis (PFGE), a

  recent progress, allows a better resolution of the frag-

  for a given voltage Schwartz and Cantor indicate that it is possible to resolve large fragments of DNA in an agarose gel

  alternation with two pulsed electric fields at about 9 Q O.

  (Schwartz D and Cantor C "Separation of Yeast Chromosome-

  As a result, the longer DNA fragments can be separated from the shorter fragments by displacement.

  forces molecules to reorient themselves

  same between impulses However, the electric field

  induced is not uniform in the gel with this technique.

  Macromolecules tend to migrate in the gel with a mobility and trajectory that depend on the place

  o the samples are loaded into the gel This distortion

  depending on the position and which is due to the lack of uniformity of the electric field, prevents comparison

track to lanes.

  Carle et al describe a simple variant of electricity

  PFGE trophoresis which is gel electrophoresis with reverse

  The field of view (FIGE) in which the alternating fields are at 1800 The polarity is inverted at intervals with a longer pulse time in the forward direction than in the opposite direction, so that a resulting migration of the forward sample is obtained This technique

  gives acceptable resolution up to 800 kilobases (kb).

  In the case of fragments larger than a certain size, however, large and small molecules of DNA tend to migrate together.

  (by ramp effect) of the voltage during the electro-

  Phoresis allows a spread of the separation range before this phenomenon occurs. However, a set of parameters suitable for the size of the molecule to be separated must be selected initially (see Carle G, Frank M. and Olson M, "Electrophoretic Separations of large DNA Molecules by Periodic Conversion of the Electric Field ",

Science 232: 65, 1986).

  Gardiner and Patterson describe transverse alternating field electrophoresis (TAFE) in which the gel is oriented vertically and the usual matrix of four electrodes is not placed in the plane of the gel but in front of and behind it (Gardiner K , Laas W. and Patterson D, "Fractionation of Large Mammalian DNA Restriction Fragments Using Vertical Pulsed-Field Gradient Gel Electrophoresis," Som Cell Molecular Genetics 12: 185,

1986).

  Chu et al indicate that the critical factor for the resolution of large DNA fragments is the maintenance of the angle between the two directions of the AC field at a value greater than 90 (Chu G, Vollrath D and Davis R, "Separation of A Homogeneous Electric Fields Homogeneous Electric Fields ", Science 234: 1582, 1986) A homogeneous electric field system locked to the contour or CHEF system uses a hexagonal matrix having pairs of electrodes set at 1200. The use of transistors for to put the electrodes at their ideal voltages ensures the locking of the field An increased resolution is obtained without simultaneous migration of the samples CHEF-DR II (BIO-RAD, Richmond, California) uses this technique and it is used very importantly for the separation of large molecules

  DNA (U.S. Patent No. 4,878,008).

  Current systems, however, require that the user manually determine and enter the parameters of the electrophoresis operation This technique is not very effective because it requires a lot of empirical tests by the user More specifically, this

  approach prevents the benefit of a knowledge base

  This invention relates to novel techniques and apparatus which provide these advantages.

  The invention relates to a system for separating

  electrophoresis of macromolecules using a computer The system maintains a knowledge base derived from calibration data obtained by separation of macromolecules, in particular DNA, of known size,

  under various planned conditions chosen for the optimization

  separations In a preferred embodiment

  the knowledge base is incorporated as constants, functions, and tables as part of the

CHEF representation program.

  During operation, the user enters the smallest and largest size of DNA that must be

  On the basis of this information, the system

  automatically reads an electrophoresis device with the optimal parameters of voltage gradient, angle,

  initial switching time, final switching time

  voltage gradient in the forward direction and voltage gradient in the opposite direction, so that the

  resolution of electrophoresis is increased.

  Other features and advantages of the invention

  tion will become more apparent from the following description,

  with reference to the accompanying drawings in which: FIG. LA is a block diagram of the system according to the invention;

  Figure 1B is a diagram of the representative program

  tation and the electrophoresis apparatus; Fig. 2 is a flow chart of the CHEF representation program; Fig. 3 is a flow chart in case 1; FIGS. 4A and 4B are a flowchart for determining the switching times for case 1; Fig. 5 is a flow chart of the mobility averaging subprogram; Fig. 6 is a flow chart illustrating case 2; Fig. 7 is a flowchart illustrating the determination of the initial switching times in case 2; Fig. 8 is a flowchart for determining the final switching times in case 2; Figs. 9A and 9B show a mobility determination flow chart in case 2; Fig. 10 is a flow chart of case 2; Figs. 1A and 11B are a flow chart of case 4; and FIG. 12 is a flowchart of the determination

  initial switching times in case 4.

  The present invention relates to methods and

  appliances that are used with an electro-

  phoresis so that the separation and purification of macromolecules, especially DNA, are improved according to

  invention, a user enters information relating to

  to the macromolecular sample that must be separated Parameters that optimize the separation are then determined Finally, the electrophoresis device is

calibrated with these parameters.

  Reference is first made to Figure 1A; the present invention may be implemented in a system such as system 100 which includes a computer assembly 150, a CHEF display assembly 150, and an electrophoresis device or apparatus 130. The computer assembly 150 further includes central processor 108, system memory 107, keyboard or input device 103, mouse 102, fixed disk 104, display monitor 101, external interface 105, printer 109 and input controller The elements of the computer set 150 are coupled to a system bus 115 which forms a link between all the elements. In a preferred embodiment, a suitably programmed IBM compatible personal computer 10 is used.

  (International Business Machines, Armonk, New York)

  valiant under "Windows" from Microsoft (Microsoft, Redmond, Washington). Referring to Figure 1B; the display assembly 120 CHEF and the electrophoresis apparatus 130 are shown in greater detail. The assembly 120 is coupled to the device 130 and can therefore directly calibrate the device 130 with optimized parameters. In a preferred embodiment, the device 120 Apparatus 130 is an electrophoresis device employing the techniques CHEF Des

  samples of macromolecules, which are placed initially

  in cavities, migrate in the gel under the action of an electric field More precisely, the samples migrate under the action of a vector electric field created

by several electrodes.

  In one embodiment, the assembly 120 is also coupled to an external interface 105, for example an RS-232 serial channel, allowing the assembly 120 to receive the output signals from the set 150 which is used.

  itself for the calibration of the electrophoric apparatus 130

  In another embodiment, the assembly 120 is coupled to its own input device, such as a keyboard 121 and / or a bar code reader 122, so that the assembly 120 can receive information directly. of the user, with different input formats, or from other sets As indicated in FIG. 1A, the assembly 120 can also be coupled to an external interface 105 and to the keyboard 121 and to the code reader.

bars 122, simultaneously.

  The installation 100 works in totally

  automatic; the user enters the sizes of the fragments

  the smallest and largest (expressed in kilobases) macromolecules (eg DNA) which are expected to be separated A knowledge base including the relationships discovered by the Applicants is given and allows the system 100 to automatically determine optimal parameters such as the forward voltage gradient, reverse voltage gradient, voltage gradient, angle, initial switching time, final switching time and duration

  of the test, as well as the best electro-

  phoresis Thus, maximum separation of macromolecules

can be realized.

  In one variant, the system 100 works in

  semi-automatic; the user can specify

  additional meters, including the type of agarose, the

  concentration of agarose, the type of buffer, the concentration

  This value is then used, along with the sizes of the DNA, for determination.

optimal settings.

  The individual parameters of the system 100 are now described in more detail. The forward voltage gradient expresses the voltage difference by

  unit of distance (V / cm) that is used during the impulse

  in the direct direction of electrophoresis Fig. The reverse voltage gradient on the other hand and the voltage difference per unit distance (V / cm) of the inverse pulse The voltage gradient is simply the difference between voltage gradients in the direction

direct and in the opposite direction.

  The switching time is the time during which the electrophoresis apparatus 130 remains in a particular state (before new switching). Thus, this is the period during which the system keeps a particular set of vectors (voltage gradients for a particular angle) The switching time varies gradually (ramp) during electrophoresis Preferably, the switching time varies in an increasing manner Preferably, the switching time varies in an increasing and non-linear manner. preferred embodiment, the switching time varies logarithmically

  In a variant, the switching time

  tion varies hyperbolically In another mode of

  the user specifies the gradual variation by determining a curve in a coordinate system, for example a Cartesian coordinate system in which X is the normalized duration of the test and Y is the normalized difference between the initial times and Final Switching Independently of the progressive dimming technique used, it is preferable to divide the switching time into initial and final switching times indicating the upper and lower limits of the switching time.

variation of the switching time.

  The angle and duration of the test retain their significance.

  usual cation The angle is the angle maintained between the alternating fields and the sample Historically, the forward or downward direction is defined as an angle equal to 0 and the reverse or upward direction corresponds to an angle of 180 Finally the duration of the test is the period during which the electrophoresis is performed by the electrophoresis apparatus 130. As a user may be familiar with a particular set of terminologies, three different types of programs are used in a preferred embodiment: 1) FIG, 2) two-state CHEF, and 3) multi-state CHEF. user by

  example for the observation of the parameters of the test.

  FIGE electrophoresis has only one pulse in the

  forward direction (angle = O) and in the opposite direction (angle = 1800).

  In the multi-state CHEF program type, forward processing occurs when the angle is 0, but there are also offsets, positive or negative. For example, 600 on one side gives a corresponding offset of 60 so the resulting load is in the forward direction The two-state CHEF is similar to the multi-state CHEF, but uses an included angle So for angles

  600 and 600, there is an included angle of 120.

  The three types of programs present

  between infrastructure and superstructures.

  All types of programs can be described by the type of CHEF multistate program This type of program can include up to eight blocks, each block

  being able to contain up to fifteen states (vectors).

  The two-state program type includes a block with two symmetric states. All values for each

  parameter in the two vectors are identical, with the exception

  ception angle The angles are symmetrical so that

their sum is zero (direct sense).

  Electrophoresis FIGE is also a special case of the multi-state CHEF technique It also includes a block with two vectors However, these vectors are not symmetrical A vector always has an angle O (direct)

  and the other an angle of 1800 (inverse).

  Under normal conditions in fully automatic mode, the following values are assumed for small DNA sizes (<2500 kb): 1) agarose type = "low EEO" (industry standard), 2) agarose concentration = 1.0% 3) buffer type = TBE (tris-borate EDTA) 4) buffer concentration = 0.5 X TBE, and) buffer temperature = 14 OC. The following normal conditions are assumed for larger DNA sizes: 1) agarose type = low EEO, 2) agarose concentration = 0.8% 3) buffer type = TAE (tris-acetate EDTA), 4 buffer concentration = 1, OX TAE, and buffer temperature = 14 OC. System 100 knowledge base is derived

  calibration data obtained by molecular separation

  DNA size of known size in various conditions

  planned for the optimization of separations.

  In a preferred embodiment, the knowledge base is in the form of constants, functions and

  tables forming part of the CHEF representation program.

  The functions were obtained by polynomial and curvilinear regressions The generated surfaces were adapted to a least-squares displacement procedure

  weighted by distance, performed on normal data

  (see Lancaster, Peter and Salkauskas, "Curve and Surface Fitting: An Introduction", Academic Press, 1974, and Mac Lain, "Drawing Contours from Arbitrary Data Points,"

  Comput J, 17: 318-324, 1974) The functions and sur-

  matching faces are used for interpolation and in

some cases for extrapolation.

  Effect of DNA size The fully automatic operation of the system 100 is divided into four different cases, the actual case being determined by the maximum and minimum DNA sizes entered by the user In a preferred embodiment, three ranges DNA sizes are determined:

  (1) 1-50 kb, (2) 50-2,200 kb and (3) more than 2,500 kb.

  Each beach has its own particular game of characteris-

  macromolecular ticks that can be exploited for

the optimization of the separation.

  For DNA fragments of 1 to 50 kb for example,

  movement in a pulsed field is a logistic function

  This is different from the larger fragments

  DNA fragments that have mobility that is linear as a function of size Fragments of different sizes do not have an identical response to fields of different intensities The relatively large fragments, for example from 30 to 50 kb, move relatively faster at 9 V / cm

  fragments of about 1 to 25 kb Different intensities

  Fields can be used for reducing the large spacing (distance between DNA bands)

  obtained with smaller bands (less than 10 kb).

  As a result, a more uniform spacing occurs over the entire range. Thus, a preferred embodiment uses a field inversion technique with different voltages for the fields in the forward direction and in the opposite direction for the fields. fractions smaller than kb so that the fragments are dispersed in a more uniform manner (relative to the pulsed fields of

identical voltages).

  The DNA between 50 and 2500 kb represents the range that was originally described by Schwartz and Cantor. Most pulsed-field gel separations are intended for

separate the DNA in this range.

  For the separation of large fragments of DNA,

  field strength must be reduced so that large molecules can not get entangled or break in the gel However, these low field strengths require extremely long test times as the angle between the two fields has an influence on DNA mobility, test times can be reduced by reducing the

the angle between the fields.

  System Operation The operation of the system 100 is now described in detail. FIG. 2 generally represents the CHEF program 200 which is a set of procedures or steps stored in a system memory 107 for instructing the processor.

  In another embodiment, the program

  gram 200 is stored in a firmware, for example in a ROM, and is coupled to the bus 115 of the

  system or directly to the assembly 120.

  At the start of program 200, step 201, the use of

  sator enters the input parameters with the keyboard 103 and / or the mouse 102; if the program 200 is directly coupled to the set 120, the parameters are preferably entered with the keypad 121 or the bar code reader 122. In automatic mode, the user only has to enter the minimum size of the DNA and the size maximum amount of DNA from the sample to be separated In non-automatic mode,

  the user also grasps the type of agarose, the concentration

  agarose, buffer type, buffer concentration and temperature Regardless of the mode, the parameters entered are stored in a table as Input Parms, loaded into memory 107 and / or on a disk.

fixed 104.

  In step 202, if the maximum size of DNA entered ("High") is less than or equal to 50 kb, the procedure goes to step 203 for the execution of case 1 Moreover, the procedure goes to step 204 If the value High is less than or equal to 2,500 kb in step 204, the procedure goes to step 205 and executes case 2.

  step 206 If the High value is lower than

  6,000 kb or more in step 206, the procedure proceeds to step 208 which determines whether the minimum DNA size ("Low") is less than or equal to 2,000 kb. If the response is positive, the subprogram executes case 4 in step 210 Otherwise, the procedure goes to case 3 of

not 209.

  However, if the High value is not less than or equal to 6,000 kb in step 206, step 207 sets all

  output values to 0 After this step or after executing

  In cases 1 to 4, the procedure proceeds to step 211 and saves all the output values in an Output Parms table. At step 212, these values are transmitted to the set 120, either directly by the program 200 or through the interface. external 105 The set 120 then calibrates

  the electrophoresis apparatus 230 with the optimum parameters

  angle, initial switching time, voltage, duration of the test, forward voltage and

voltage in the opposite direction.

Individual cases

  The individual cases of the 200 representation program

  Figure 3 shows the steps of Case 1, that is, when the High value is less than or equal to 50 kb At step 301, the gel electrophoresis technique. In step 302, the forward voltage gradient is set to 9 V / cm At step 303, the reverse voltage gradient is set to 6 V / cm At step 304, the reverse voltage gradient is set to 9 V / cm. angle is set to 1800 and the travel distance is set to 10 cm in step 305,

  the switching times are determined.

  Figures 4A and 4B show a flowchart

  illustrating the steps involved in determining the commu-

  In step 401, if the value Low is less than kb, step 402 sets the initial switching time to 1.2 s. In addition, the procedure proceeds to step 403 which calculates the initial switching time with the following relation: function 1 L) Y = 0.0 + 0.015797 X + 0, 001365 x 2 X being the Low value and Y the switching time in seconds At step 404, if the initial switching time is between 0.0 and 0.05 s, the initial switching time is set to 0.05 S in step 405 Otherwise, step 405 is skipped In step 406, the final switching time is determined according to the following relation (function l U) Y = 0.0 + 0.009166 X + 0.000137 x 2 X being the High value and Y the switching time in seconds The Y result is multiplied by 1.15, an empirically determined adjustment factor. 407, if the final switching time is between 0.0 and 0.05 s, step 408 sets this final time to 0.05 s Otherwise, step 408 is skipped In step 409, if the initial switching time is greater than the final switching time, step 410 reverses the initial and final switching times (replacing one by the other). If not, step 410 is skipped. The subprogram

returns and finishes step 305.

  At step 306, if the Low value is between 1 and 6 kb or is equal to either of these values, at step 308, the mobility (in cm / h) is determined from the following relationship (function 2) Y = 0.65426181 0, 0838514428 X + 0.0060879216 x 2 X being the Low value However, if this value is not in the range of step 306, step 307 determines the

  mobility (cm / h) in Table 1 (Appendix) and the sub-component

  training program of the average mobility.

  FIG. 5 shows the steps of the mobility averaging routine At step 501, the switching time range is determined from the initial and final switching times At step 502, the number N of iterations is determined by dividing the switching time range by 5 and rounding to the nearest integer At step 503, the switching delta time is determined by dividing the switching time range by N at step 504, the following constants are determined: sum = 0.0, i = 0, current switching time = initial switching time At step 505, i is tested for the counting of N If i is not less than or equal to N + 1, the sub - program divides the sum by N in step 506 and stops then returns to step 307 Otherwise, the subroutine continues at 507 to compute a mobility from table 1 or table 2 (appendix) by use Low value as the DNA size value, the time current switching time as switching time, and by bilinear interpolation At step 508, the mobility is added to the sum At step 509, the current switching time advances to step 510, i

  progresses, and the procedure loops on step 505.

  After the end of the mobility averaging training routine at step 307, the time of the test is

  determined in step 309 by dividing the distance of movement

  Mobility by multiplication by a calibration factor Slight variations in any one of several factors, eg water quality and laboratory technique, can significantly affect the functioning of the system. variations are minimized, the user adjusts the calibration factor based on his experience of the system The output signal of the program 200 is adjusted accordingly.

  subroutine of case 1 ends with return to step 203.

  Figure 6 illustrates the steps of Case 2 At Step

  601, the angle is set to 1200, and the distance of

  In step 602, the voltage gradient is set to 6 V / cm At step 603, the initial switching times for case 2 are determined. FIG. 7 shows the steps executed in step 701, if the value Low is less than 25 kb, at step 702, the initial switching time is determined with the following relation (function 22) Y = 0, 0 + 0.072277 X 0.008314 x 2 + 0.000581 X 3 X being the Low value and Y the switching time The procedure then jumps to step 706 However, if the Low value is not less than 24 kb in step 701, step 703, the Low value is examined so that the fact that it is less than 50 kb is determined. If the answer is

  positive step 704 determines the initial switching time

  using the following relation (function 21 U) Y = 0,0701979 + 0,074324119 X + 0,000007336094 x 2 X being the value Low and Y the switching time The subroutine continues in step 706 However if the value Low is not less than 50 kb at step 707, step 705 determines the initial switching time using the following relation (function 21 L) Y = 0.95604 + 0.16292974 X 0 , 00000574226 x 2 X being the Low value and Y the switching time At step 706, if the initial time is between 0.0 and 0.05 s, step 707 reverses the values of the initial time and the final switching time. The subroutine ends, and

  concludes step 603 of the calling subroutine.

  At step 604, the final switching time is

  Figure 8 shows the steps of this method.

  At step 801, if the value Hig H is less than 80 kb, step 802 determines the final switching time with the following relation (function 21 U) Y = 0.0701979 + 0.074324119 X + 0.000007336094 x 2

  X being the High value and Y the switching time.

  However, if the High value is less than 80 kb in step 801, the procedure proceeds to step 803 which calculates the final switching time with the following relation (function 20) Y = 0.0 + 0.007605 x + 0.000686 x 2

  X being the High value and Y the switching time (in s).

  The procedure moves to step 804 or the final switch time

  is multiplied by 1.15 In step 805, if the final time is between 0.0 and 0.05 s, the step 806 inverts the initial and final time values The procedure ends

  and concludes step 804 of the calling routine.

  In step 605, if the initial switching time is greater than the final time, step 606 reverses the initial and final time values. Otherwise, step 606 is

  jumped At step 607, the mobility of case 2 is determined.

  FIG. 9 represents the process implemented at step 607 At step 901, if the value Low is less than kb, step 902 determines the mobility (in cm / h) using the following relation (function 23)

Y = 1.0124518X-0.4094122386

  X being the value Low and Y the mobility In step 903, if the value Low is less than 169 kb and the final switching time is less than 110, 0, the mobility (cm / h) is determined in step 904 with the table 2 (appendix), the initial and final switching times, the Low value constituting the size of the DNA and the subprogram of

  training of the mobility average (Figure 5).

  hard then ends with a return.

  However, if step 904 gives a negative response, the procedure proceeds to step 905 which determines whether the Low value is less than 2,500 kb. If the response is positive, step 906 determines the initial switch time to see if it is between 110.0 and 300.0 s If the response is positive, step 907 determines the mobility (cm / h) using Table 3 (appendix), the Low value as DNA size, and linear interpolation Then, the procedure ends and concludes step 607 of the calling routine. However, if the known size of DNA is not less than 2,500 kb at step 905, steps 906 and 907 are skipped. In addition, if step 906 gives a negative response, step 907 is skipped at step 908, the value Low is read to determine if it is less than 1,169 kb If the answer is positive, step 909 determines whether the initial switching time is less than 110,0 S and the final time greater than 110,0 s, and the procedure goes to step 910

  to calculate mobility using Table 2 (

  dice), initial and final switching times,

  Low value as DNA size and the procedure of forma-

  the average of the mobility The subprogramme

then concludes with a return.

  However, if the Low value is not less than 1,169 kb in step 908, steps 909 and 910 are skipped. In addition, if step 909 gives a negative response, step 910 is skipped. program gets to step 910 to which it puts all the output values to O

before concluding at step 607.

  At step 608, the test time is set to a value equal to the calibration factor multiplied by the travel distance and divided by the mobility.

  2 ends and ends step 205.

  FIG. 10 represents the steps of the procedure of the case 3 which is called at step 209 (FIG. 2) At step 1001, the angle is set at 1060 At step 1002, the voltage gradient is set at 2 V / cm at step 1003, the initial switching time is determined with the following equation (function 50)

Y = 8.6 + 0.0053 X

  X being the value High multiplied by 60 and Y the time of

  At step 1004, if the initial switching time

  The time between 0.0 and 0.05 s, in step 1005, the initial time is set to 0.05 s Otherwise, step 1005 is skipped. At step 1006, the final switching time is set to one. value equal to the initial time In step 1007, the mobility (cm / h) is determined using the following relation: (function 51) Y = 0.047 + 0.00000251 X 1.56 10-9 x 2 X being Low and Y the mobility In step 1008, the travel distance is set to 3 cm At step 1009, the travel time is set to the calibration factor multiplied by the travel distance and divided by the mobility step 3 is

finish and conclude the step 209.

  FIGS. 11A and 11B represent the steps of case 4 called at step 210 (FIG. 2) At step 1101, the angle is set at 1060 At step 1102, the voltage gradient is set at 2 V / cm At step 1103 , the travel time is set to the calibration factor multiplied by 72 h At step 1104, the initial switching time is set to 20 min At step 1105, the final switching time is determined with the following relation (function 50)

Y = 8.6 + 0.0053 X

  X being the value High multiplied by 60 and H the mobility.

  In step 1106, the final time is multiplied by 1.15 in step 1107, if the final time is between 0.0 and 0.05 s, the

  1108 sets the final time to 0.05 s In the case of

  milk, step 1108 is skipped In step 1109, the values of

  Block 1 output are saved.

  In steps 1110 and 1111, the angle is set to 1200 and the voltage gradient to 6 V / cm At step 1112, the final switching time is set to 3 min. At step 1113, the times

  initial switching are determined.

  FIG. 12 illustrates the method for determining the initial switching times for case 4 At step 1201, if the Low value is less than 24 kb, step 1202 determines the initial switching time with the following relation: (function 22) Y = 0.0 + 0.072277 X 0.008314 x 2 + 0.000581 X 3 X being the Low value and Y the switching time The subroutine continues at step 1206 However, if the Low value is not lower at 24 kb in step 1201, the procedure proceeds to step 1203 which determines if the Low value is less than 50 kb When the answer is positive, step 1204 determines the initial switching time with the following relation (function 21 U) Y = 0.0701979 + 0.074324119 X + 0.000007336094 x 2 X being the Low value and Y the switching time The procedure continues in step 1206 However, if the Low value is not less than 50 kb in step 1203, the initial switching time is determined in step 1205 by using the following 21 L) Y = 0.95604 + 0.111662974 X 0, 00000574226 x 2

  X is the Low value and Y is the switching time.

  The procedure continues in step 1206 which determines whether the initial switching time is between 0.0 and 0.05 s If the response is positive, step 1207 makes the initial time equal to 0.05 s Otherwise, step 1207 is skipped Step 1208 determines whether the initial switching time is greater than the final time If the response is positive, step 1209 reverses the values of the initial and final switching times.

conclude and end step 1113.

  In step 1114, the mobility (cm / h) is determined in the same way as for case 2 (FIG. 9). In step 1115, the travel distance is set to 4 cm. In step 1116, the travel time is determined by multiplication of the calibration factor by the distance of travel and division by mobility The procedure concludes with a return. appendix

Table 1

  The first column (X) gives the minimum size

DNA (Low) (in kilobases).

  The second column (Y) gives the switching time

* (in seconds).

  The third column (Z) gives the mobility of elec-

  trophorèse (in centimeters per hour).

Data

X Y Z

8.14 0.2 0.273

8.14 0.5 0.338

8.14 0.8 0.384

, 18 0.2 0.245

, 18 0.5 0.315

, 18 0.8 0.362

12.20 0.2 0.217

12.20 0.8 0.351

, 00 0.2 0.180

, 00 0.5 0.281

, 00 0.8 0.348

17.00 0.2 0.157

17.00 0.5 0.262

17.00 0.8 0.339

19.40 0.2 0.138

19.40 0.5 0.236

19.40 0.8 0.332

22.60 0.3 0.177

22.60 0.5 0.216

22.60 0.8 0.318

24.80 24.80 24.80 29.90

29,90

29.90 33.50 33.50 33.50

38.40

  38.40 38.40 48.50 48.50 The first column (X) is the minimum DNA size (Low) (in kilobases). second column (Y) is the switching time

(in seconds)

  The third column (Z) is the mobility of

  trophorèse. Suitable surface XYZ 8,1400 e + 000 8,1400 e + 000 8,1400 e + 000 8,1400 e + 000 8,1400 e + 000 8,1400 e + 000 8,1400 e + 000 8,1400 e + 000 8,1400 e + 000 8, 1400 e + 000 8,1400 e + 000 2,0000 e-001 2,3000 e-001 2,6000 e-001 2,9000 e-001 3,2000 e-001 3,5000 e-001 3,8000 e-001 4.1000 e-001 4.4000 e-001 4.7000 e-001, 0000 e-001 2.7129 e-001 2.8031 e-001 2, 8722 e-001 2.9268 e-001 2.9759 e-001 3.0269 e-001 3.0842 e-001 3.1468 e-001 3.2185 e-001 3.2911 e-001 3.3611 e-001 0.3 0.5 0.8 0, 3 0.5 0.8 0.3 0, 5 0.8 0.3 0.5 0.8 0.5 0.8 0.158 0.191 0.306 0.128 0.177 0.274 0.144 0, 156 0.245 0.105 0.126 0.210 0.095 0.146 8.1400 e + 000 8,1400 e + 000 8, 1400 e + 000 8,1400 e + 000 8,1400 e + 000 8,1400 e + 000 8, I 400 e + 000 8, 1400 e + 000 8,1400 e + 000 8,1400 e + 000 1, 0158 e + 001 1.0158 e + 001 1, 0158 e + 001 1.0158 e + 001 1.0158 e + 001 1.0158 e + 001 1.0158 e + 001 1, 0158 e + 001 1.0158 e + 001 1.0158 e + 001 1, 0158 e + 001 1.0158 e + 001 1, 0158 e + 001 1.0158 e + 001 1.0158 e + 001 1.0158 e + 001 1.0158 e + 001 1, 0158 e + 001 1.0158 e + 001 1.0158 e + 001 1.0158 e + 001 1.2176 e + 001 1, 2176 e + 001 1.2176 e + 001 1.2176 e + 001, 3000 e-001, 6000 e-001, 9000 e- 001 6,2000 e-001 6,5000 e-001 6,8000 e-001 7,1 OO Oe-001 7,4000 e-001 7, 7000 e-001 8, OOO Oe-001 2, OO Oe-001 2.3000 e-001 2.6000 e-001 2.9000 e-001 3.2000 e-001 3.5000 e-001 3.8000 e-001 4.1000 e-001 4.4000 e-001 4, 7000 e-001, OOO Oe-001, 3000 e-001, 6000 e-001, 9000 e-001 6,2000 e-001 6,5000 e-001 6,8000 e-001 7,10 l OO Oe-001 7,4000 e-001 7,7000 e-001 8, OOO Oe-OO 1 2.0000 e-001 2.3000 e-001 2.6000 e-001 2.9000 e-001 3.4220 e-001 3 , 4727 e-001 3.5142 e-001 3.5486 e-001 3.5794 e-001 3.6108 e-001 3, 6467 e-001 3.6903 e-001 3.7423 e-001 3.7958 e-001 2,4379 e-001 2,5399 e-001 2,6236 e-001 2,6923 e-001 2,7539 e-001 2,8155 e-001 2,8814 e-001 2, 9523 e 001 3,0264 e-001 3,1004 e-001 3,1720 e-001 3,2401 e-001 3,3004 e-001 3,3519 e-001 3,3968 e-001 3,4383 e-001 3 , 4795 e-001 3.5227 e-001 3, 5690 e-001 3.6177 e-001 3.6691 e-001 2.117 e-001 2.2861 e-001 2 , 3852 e- 001 2.4691 e-001 1.2176 e + 001 1.2176 e + 001 1.2176 e + 001 1.2176 e + 001 1.2176 e + 001 1.2176 e + 001 1.2176 e + 001 1.2176 e + 001 1.2176 e + 001 1.2176 e + 001 1.2176 e + 001 1.2176 e + 001 1.2176 e + 001 1.2176 e + 001 1.2176 e + 001 1.2176 e + 001 1.2176 e + 001 1.4194 e + 001 1.4194 e + 001 1.4194 e + 001 1.4194 e + 001 1.4194 e + 001 1.4194 e + 001 1.4194 e + 001 1.4194 e + 001 1.4194 e + 001 1.4194 e + 001 1.4194 e + 001 1.4194 e + 001 1.4194 e + 001 1.4194 e + 001 1.4194 e + 001 1.4194 e + 001 1.4194 e + 001 1.4194 e + 001 1.4194 e + 001 3.2000 e-001 3.5000 e-001 3,8000 e-001 4, 1000 e-001 4,4000 e-001 4,7000 e-001, OOO Oe-001, 3000 e-001, 6000 e-001, 9000 e-001 6.2000 e-001 6,5000 e-001 6,8000 e-001 7,1000 e-001 7, 4000 e-001 7,7000 e-001 8,0000 e-001 2, OOO Oe-001 l 2,3000 e -001 2.6000 e-001 2.9000 e-001 3.2000 e-001 3.5000 e-001 3.8000 e-001 4.1000 e-001 4.000 e-001 4.7000 e-001 , 000 Oe-001, 3000 e-001, 6000 e-001, 9000 e-001 6,2000 e-001 6,5000 e-001 6,8000 e-001 7,1000 e-001 7,4000 e-001 2 5435 e-001 2.6154 e-001 2.683 e-001 2.7663 e-001 2.8453 e-001 2.9237 e-001 2, 9990 e-001 3.0694 e-001 3.1339 e -001 3,1930 e-001 3,2481 e-001 3,3013 e-001 3,3540 e-001 3,4071 e-001 3,4605 e-001 3,5132 e-001 3,5688 e-001 1, 9194 e-001 2.0479 e-001 2.1642 e-001 2.2635 e-001 2.394 e-001 2.4292 e-001 2.5084 e-001 2.5893 e-001 2, 6725 e-001 2,7564 e-001 2,8362 e-001 2, 9064 e-001 2,9722 e-001 3,0374 e-001 3,1026 e-001 3,1678 e-001 3,2333 e - 001 3,2989 e-001 3,3643 e-00 @ 1,4194 e + 001 1,4194 e + 001 1,6212 e + 001 1,6212 e + 001 1,6212 e + 001 1,6212 e + 001 1,6212 e + 001 1.6212 e + 001 1.6212 e + 001 1.6212 e + 001 1.6212 e + 001 1.6212 e + 001 1.6212 e + 001 1, 6212 e + 001 1.6212 e + 001 1.6212 e + 001 1.6212 e + 001 1.6212 e + 001 1, 6212 e + 001 1.6212 e + 001 1.6212 e + 001 1.6212 e + 001 1.6212 e + 001 1, 8230 e + 001 1.8230 e + 001 1.8230 e + 001 1.8230 e + 001 1.8230 e + 001 1, 8230 e + 001 1.8230 e + 001 1.8230 e + 001 1.8230 e + 001 1.8230 e + 001 1, 8230 e + 001 1.8230 e + 001 7.7000 e-001 8, OOO Oe -001 2, OO O Oe-001 2, 3000 e-001 2.6000 e-001 2.9000 e-001 3.2000 e-001 3.5000 e-001 3.8000 e-001 4.1000 e-001 4.4000 e -001 4,7000 e-001, 0000 e-001, 3000 e-001, 6000 e-001, 9000 e-001 6,2000 e-001 6,5000 e-001 6,8000 e-001 7,10 l OO Oe- 001 7,4000 e-001 7,7000 e-001 8,0000 e-001 2,0000 e-001 2,3000 e-001 2, 6000 e-001 2,9000 e-001 3,2000 e -001 3.5000 e-001 3.8000 e-001 4.1000 e-001 4.4000 e-001 4.7000 e-001, 0000 e-001, 3000 e-001 3.4295 e-001 3, 4960 e-001 1.6937 e-001 1.8355 e-001 1.9689 e-001 2.0824 e-001 2.174 e-001 2.2594 e-001 2.3394 e-001 2.4202 e -001 2,5029 e-001 2,5870 e-001 2, 6677 e-001 2,7406 e-001 2,8112 e-001 2,8842 e-001 2,9598 e-001 3,0371 e- 001 3,1154 e-001 3,1938 e-001 3,2717 e-001 3,3486 e-001 3,4240 e-001 1, 5044 e-001 1,6570 e-001 1,8059 e-001 1, 9314 e-001 2.0283 e-001 2.1081 e-001 2.1834 e-001 2.2594 e-001 2.3368 e-001 2.4146 e-001 2.4922 e-001 2. 5704 e -001 2.6504 e-001 2.7333 e-001 2.8195 e-001 2.9082 e-001 2.9982 e-001 3.0883 e-001 3.1773 e-001 3.2641 e-001 3,347 2 e-001 1.8230 e + 001 1, 8230 e + 001 1.8230 e + 001 1.8230 e + 001 1.8230 e + 001 1.8230 e + 001 1, 8230 e + 001 1.8230 e + 001 1.8230 e + 001, 6000 e-001, 9000 e-001 6,2000 e-001 6,5000 e-001 6,8000 e-001 7,10 O Oe-001 7,4000 e-001 7,7000 e-001 8, 00 00 e-001 2.00 00 e-001 2.3000 e-001 2.6000 e-001 2.9000 e-001 3.2000 e-001 3.5000 e-001 3.000 e-001 4.10 O Oe-001 4.4000 e-001 4.7000 e-001, 0000 e-001, 3000 e-001, 6000 e-001, 9000 e-001 6.2000 e-001 6.5000 e-001 6, 8000 e-001 7.10 O Oe-001 7.4000 e-001 7.7000 e-001 8.0000 e-001 1, 3526 e-001 1.5140 e-001 1.6750 e-001 1, 8085 e-001 1,9028 e-001 1,9738 e-001 2,0406 e-001 2,1092 e-001 2,1793 e-001 2,2507 e-001 2,3254 e-001 2, 4068 e -001 2,4943 e-001 2,5864 e-001 2,6823 e-001 2,7808 e-001 2,8806 e-001 2,9805 e-001 3,0790 e-001 3,1746 e-001 3,2646 e-001 2,0248 e + 001 2, 0248 e + 001 2,0248 e + 001 2,0248 e + 001 2,0248 e + 001 2,0248 e + 001 2, 0248 e + 001 2,0248 e + 001 2,0248 e + 001 2.0248 e + 001 2.0248 e + 001 2, 0248 e + 001 2.0248 e + 001 2.0248 e + 001 2.0248 e + 001 2.0248 e + 001 2, 0248 e + 001 2.0248 e + 001 2.0248 e + 001 2.0248 e + 001 2.0248 e + 001 1, 2491 e-001 1.4088 e-001 1 , 5667 e-001 1,6970 e-001 1,7860 e-001 2,2266 e + 001 2,2266 e + 001 2,2266 e + 001 2,2266 e + 001 2,2266 e + 001 2,00 00 e- 001 2 , 3000 e-001 2.6000 e-001 2.9000 e-001 3.2000 e-001 2.2266 e + 001 2, 2266 e + 001 2.2266 e + 001 2.2266 e + 001 2.2266 e + 001 2, 2266 e + 001 2, 2266 e + 001 2,2266 e + 001 2,2266 e + 001 2,2266 e + 001 2,2266 e + 001 2, 2266 e + 001 2,2266 e + 001 2,2266 e + 001 2,2266 e + 001 2, 2266 e + 001 2, 4284 e + 001 2.4284 e + 001 2.4284 e + 001 2.4284 e + 001 2.4284 e + 001 2, 4284 e + 001 2.4284 e + 001 2.4284 e + 001 2.4284 e + 001 2, 4284 e + 001 2, 4284 e + 001 2.4284 e + 001 2.4284 e + 001 2.4284 e + 001 2.4284 e + 001 2, 4284 e + 001 2.4284 e + 001 2.4284 e + 001 2.4284 e + 001 2, 4284 e + 001 3, 5000 e-001 3,8000 e-001 4,1000 e-001 4,4000 e-001 4,7000 e-001, 0000 e-001, 3000 e-001, 6000 e-001, 9000 e-001 6,2000 e-001 6,5000 e-001 6,8000 e-001 7,10 l O Oe-001 7,4000 e-0 01 7.7000 e-001 8.0000 e-001 2.0000 e-001 2.3000 e-001 2.6000 e-001 2.9000 e-001 3.2000 e-001 3.5000 e-001 3 , 8000 e-001 4.10 l OO Oe-001 4.4000 e-001 4.7000 e-001, 0000 e-001, 3000 e-001, 6000 e-001, 9000 e-001 6.2000 e- 001 6,5000 e-001 6,8000 e-001 7, 1000 e-001 7,4000 e-001 7,7000 e-001 1,8443 e-001 1,9091 e-001 1,9707 e- 001 2 , 0335 e-001 2.0988 e-001 2.117 e-001 2.254 e-001 2.3461 e-001 2, 4457 e-001 2.5490 e-001 2.6547 e-001 2.7617 e-001 2,8689 e-001 2,9749 e-001 3,0776 e-001 3,1732 e-001 1,1872 e-001 1,3323 e-001 1,4694 e-001 1, 5821 e 001 1.6653 e-001 1.7291 e-001 1.7873 e-001 1.8449 e-001 1.9016 e-001 1.9584 e-001 2.0242 e-001 2, 11 O Oe-001 2,2091 e-001 2,3134 e-001 2, 4208 e-001 2,5303 e-001 2,6411 e-001 2,7523 e-001 2,8625 e-001 2,9696 e-001 2, 4284 e + 001 2.6302 e + 001 2.6302 e + 001 2.6302 e + 001 2.6302 e + 001 2.6302 e + 001 2.6302 e + 001 2.6302 e + 001 2.6302 e + 001 2.6302 e + 001 2, 6302 e + 001 2.6302 e + 001 2.6302 e + 001 2.6302 e + 001 2.6302 e + 001 2.6302 e + 001 2.6302 e + 001 2,6302 e + 001 2,6302 e + 001 2,6302 e + 001 2,6 * 02 e + 001 2,6302 e + 001 2,8320 e + 001 2,8320 e + 001 2,8320 e + 001 2,8320 e + 001 2 , 8320 e + 001 2,8320 e + 001 2,8320 e + 001 2,8320 e + 001 2,8320 e + 001 2,8320 e + 001 2,8320 e + 001 2,8320 e + 001 2,8320 e + 001 8, OO Oe-001 2. OO Oe-001 2.3000 e-001 2.6000 e-001 2.9000 e-001 3, 2000 e-001 3.5000 e-001 3.8000 e-001 4.10 l OO Oe-001 4,4000 e-001 4, 7000 e-001, OO Oe-001, 3000 e-001, 6000 e-001, 9000 e-001 6,2000 e-001 6,5000 e-001 6,8000 e-001 7.10 l OO Oe-001 7.4000 e-001 7.7000 e-001 8, 0000 e-001 2.0000 e-001 2.3000 e-001 2.6000 e-001 2.9000 e-001 3,2000 e- 001 3,5000 e-001 3,8000 e-001 4,1000 e-001 4,4000 e-001 4,7000 e-001, 0000 e-001, 3000 e-001, 6000 e- 001 3.0691 e-001 1.1480 e-001 1.2723 e-001 1.3835 e-001 1.4760 e-001 1.5520 e-001 1.6171 e-001 1.6764 e-001 1 , 7329 e-001 1,7881 e-001 1,8440 e-001 1,9087 e-001 1,9924 e-001 2,0889 e-001 2,1913 e-001 2,2979 e-001 2,4074 e-001 2.5188 e-001 2.6306 e-001 2, 7413 e-001 2.8489 e-001 2 , 9497 e-001 1,1212 e-001 1,2265 e-001 1,3138 e-001 1,3849 e-001 1,4532 e-001 1,5190 e-001 1,5779 e-001 1,6332 e-001 1, 6896 e-001 1.7516 e-001 1.8209 e-001 1.8965 e-001 1.9819 e-001 2.8320 e + 001 2.8320 e + 001 2.8320 e + 001 2, 8320 e + 001 2,8320 e + 001 2,8320 e + 001 2,8320 e + 001 2,8320 e + 001 3,0338 e + 001 3,0338 e + 001 3,0338 e + 001 3,0338 e + 001 3,0338 e + 001 3, 0338 e + 001 3.0338 e + 001 3.0338 e + 001 3.0338 e + 001 3.0338 e + 001 3.0338 e + 001 3.0338 e + 001 3.0338 e + 001 3.0338 e + 001 3.0338 e + 001 3, 0338 e + 001 3.0338 e + 001 3.0338 e + 001 3.0338 e + 001 3.0338 e + 001 3.2356 e + 001 3.2356 e + 001 3.2356 e + 001 3.2356 e + 001 3.2356 e + 001 3, 2356 e + 001 3.2356 e + 001, 9000 e-001 6,2000 e-001 6,5000 e-001 6,8000 e-001 7,1000 e-001 7,4000 e-001 7,7000 e-001 8 , 0000 e-001 2.3000 e-001 2.6000 e-001 2.9000 e-001 3.2000 e-001 3.000 e-001 3.000 e-001 4.1000 e-001 4.4000 e-001 4,7000 e-001, 0000 e-001, 3000 e-001, 6000 e-001, 9000 e-001 6,2000 e-001 6,5000 e-001 6,8000 e-001 7,10 l OO Oe-001 7 , 4000 e-001 7.7000 e-001 8, OO Oe-001 2.0000 e-001 2.3000 e-001 2.6000 e-001 2.9000 e-001 3.2000 e-001 3.5000 e-001 3, 8000 e-001 2.0770 e-001 2.1779 e-001 2.2859 e-001 2.3952 e-001 2.5052 e-001 2.6142 e-001 2.7204 e- 001 2,8211 e-001 1,1966 e-001 1,2721 e-001 1, 3283 e-001 1,3844 e-001 1,4414 e-001 1,4922 e-001 1,5416 e-001 1 , 5954 e- 001 1.6574 e-001 1.7260 e-001 1.7974 e-001 1.8762 e-001 1.9651 e-001 2, 0623 e-001 2.161 e-001 2.2707 e-001 2,3774 e-001 2,4835 e-001 2,5880 e-001 2,6883 e-001 1,0830 e-001 1,1739 e-001 1,2552 e-001 1,3183 e- 001 1, 3551 e-001 1.3826 e-001 1.4147 e-001 3.2356 e + 001 3.2356 e + 001 3.2356 e + 001 3.2356 e + 001 3.2356 e + 001 3.2356 e + 001 3 , 2356 e + 001 3.2356 e + 001 3.2356 e + 001 3.2356 e + 001 3.2356 e + 001 3.2356 e + 001 3.2356 e + 001 3.2356 e + 001 3.4374 e + 001 3.4374 e + 001 3 , 4374 e + 001 3,4374 e + 001 3,4374 e + 001 3,4374 e + 001 3,4374 e + 001 3,4374 e + 001 3,4374 e + 001 3,4374 e + 001 3,4374 e + 001 3,4374 e + 001 3 , 4374 e + 001 3,4374 e + 001 3,4374 e + 001 3,4 374 e + 001 3.4374 e + 001 3.4374 e + 001 3.4374 e + 001 3.4374 e + 001 3.4374 e + 001 4.10 l OO Oe-001 4.4000 e-001 4.7000 e- 001, 0000 e-001, 3000 e-001, 6000 e-001, 9000 e-001 6,2000 e-001 6,5000 e-001 6,8000 e-001 7,10 l OO Oe-001 7,4000 e-001 7.7000 e-001 8, OOO Oe-001 2.0000 e-001 2.3000 e-001 2.6000 e-001 2.9000 e-001 3.2000 e-001 3.000 e-001 3 8000 e-001 4.1000 e-001 4.4000 e-001 4.7000 e-001, 0000 e-001, 3000 e-001, 6000 e-001, 9000 e-001 6.2000 e-001 6, 5000 e-001 6,8000 e-001 7,1000 e-001 7,4000 e-001 7,7000 e-001 8,0000 e-001 1,4534 e-001 1, 5000 e-001 1,5559 e -001 1.6197 e-001 1.6897 e-001 1.7670 e-001 1.8526 e-001 1.9458 e-001 2.0443 e-001 2.1446 e-001 2.2478 e-001 2.3890 e-001 2, 4477 e-001 2.5437 e-001 1.0519 e-001 1.1328 e-001 1.2088 e-001 1.2710 e-001 1.3004 e-001 1, 3144 e-001 1.3352 e-001 1.3648 e-001 1.4031 e-001 1, 4517 e-001 1.5108 e-001 1.5789 e-001 1.6554 e-001 1.7396 e -001 1,8297 e-001 1,9241 e-001 2,0207 e-001 2,1176 e-001 2,2134 e-001 2,3072 e-001 2, 4001 e-0 01 3,6392 e + 001 3,6392 e + 001 3,6392 e + 001 3,6392 e + 001 3, 6392 e + 001 3,6392 e + 001 3,6392 e + 001 3,6392 e + 001 3,6392 e + 001 3, 6392 e + 001 3,6392 e + 001 3,6392 e + 001 3,6392 e + 001 3,6392 e + 001 3, 6392 e + 001 3,6392 e + 001 3,6392 e + 001 3,6392 e + 001 3,6392 e + 001 3, 6392 e + 001 3,6392 e + 001 3,8410 e + 001 3,8410 e + 001 3,8410 e + 001 3, 8410 e + 001 3,8410 e + 001 3,8410 e + 001 3,8410 e + 001 3,8410 e + 001 3, 8410 e + 001 3,8410 e + 001 3,8410 e + 001 3,8410 e + 001 3,8410 e + 001 3, 8410 e + 001 3,8410 e + 001 2, OOO Oe-001 2,3000 e-001 2,6000 e-001 2 , 9000 e-001 3.2000 e-001 3.5000 e-001 3.8000 e-001 4.10 l OO Oe-001 4.4000 e-001 4.7000 e-001, OOO Oe-001, 3000 e-001, 6000 e-001, 9000 e-001 6,2000 e-001 6,5000 e-001 6,8000 e-001 7,10 l OO Oe-001 7,4000 e-001 7,7000 e- 001 8,0000 e-001 2,000 Oe-001 2,3000 e-001 2,6000 e-001 2,9000 e-001 3, 2000 e-001 3,5000 e-001 3,8000 e-001 4, 10 O Oe-001 4.4000 e-001 4.7000 e-001, 0000 e-001, 3000 e-001, 6000 e-001, 9000 e-001 6.2000 e-001 1, 0050 e-0 01 1,0656 e-001 1,1172 e-001 1,1606 e-001 1,1962 e-001 1,2243 e-001 1,2490 e-001 1,2749 e-001 1,3057 e-001 1 , 3449 e-001 1.3975 e-001 1, 4654 e-001 1.5436 e-001 1.6275 e-001 1.7151 e-001 1.8054 e-001 1.8970 e-001 1.9884 e-001 2.0783 e-001 2.1667 e-001 2.251 e-001 9.5101 e-002 9, 9338 e-002 1.0269 e-001 1.0589 e-001 1.0962 e- 001 1,1328 e-001 1,1622 e-001 1,1875 e-001 1,2134 e-001 1,2447 e-001 1,2905 e-001 1,3577 e-001 1, 4370 e-001 1 , 5198 e-001 1.6040 e-001 o 00-g Os O 'Ioo-a 990 o'I ZOO-e T'86 The 6 ZOO-e S And T' 6

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TOO + 9 OTT'8 '8

TOO + 9 OTT'8 '8

  1 OO + eolt T'8 '8 ST OT S ú 990 z 9 Z 00-809 The L 10-a 008 o T O + 8 ef 9 z

TOO-865 ZL'T

  TOO-e Z Sg 9 'T TOO-8 f 895' T TOO-e Z 96 'T' T 00-898 qz 'I T 00-88 f 9' T

-8980 1 '

T 00-88 fzl 'I T 00-a Z 8 IT' T

TO-886 ZIT'T

TOO-T 8 LO 'T

-a 890 'T Z 00-88 Lg 6' 6

ZOO-89 ZU 9 '6

ZOO-a LOL '6 E 00-a 8 IEI'6

ZOO-LZT 88 '8

l 00-8 i 9 9 '8 Z 00-e Z 6 "' 8

Z 00-885 TZ'8

Z 00-8 Z O f O '8

T 100-8998 'T

OO 0-88 úLL 'I

  T 00-86 e 69 'I T 00-a 919 T' I TOO-q 1 Its' I T 00-T 8169 f'T TOO-8 f 86 ú'T

T 100-868 Z 'T

-8609 Z'T

T 00-8696 T'T

T 00-a ZTIT 'I

T 100-88960 'T

-a 6190 'T

T 00-80000 '8

-000 The IL

-OO'00 IL

TOO-OOOI'L

-80008 '9

TO 0-a OO-'9

-OOOZ'9

-OO 06 'S

OO 0-0009 '9

-OOE'S

  oooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooo

OO 0-0008 '8

l O 0-ao O Og O '

-00 Z'8

-0006 'Z

Oo-a 0009 'z-a OOOu'z

-80000 '

TO 0-O 00 '8

  -a OOOL'L TOO-a OO Of'L TO 0-a OO Ol'L

-80008 '9

-OO 0005 '9

-OOOZ'9

-90006 'S

TOO-80009 '95

0 0 0 0 0 0 0 0 0 0 0

TOO-OOO O 'S

-80000 'S

  TO 0-a OOOL'f T Oo-aooo 00 'T 00 + qf 19 fifi'fi

TOO + 1

  -'8 f 95 f 9 "f TOO + ef 19" fl 'Too + Gpg, p J i TOO + 8 f 19 lfl fi TOO + afi 9 "ii: i TOO + 8 fi 9 fifi' fi T 00 + 91 i 9 flf Jfi TO O + 8 fi 9 fi 'f T OO + efig Tif' f T 00 + f 9 fif 'fi, To O + 89 flfv' f To O + e 9 fif, 'f 0 Oo + a 9 fifp' f 00 o + 89 fifi fi 00 o + 89 ifi Ppi T 00 o 89 fifi 'fi T 00 + 89 fifi J fi 1 OT S loo + azg-pg J, zoo-e Tog LIL ú 90 z 9 Z 7, 9176 e-002 8,0843 e-002 8,2690 e-002 8,4756 e-002 8,7101 e-002 8, 9849 e-002 9,3127 e-002 9,6985 e-002 1,0136 e -001 1,0612 e-001 1,1109 e-001 1,1613e-001 1,2120 e-001 1,2639 e-001 1,3188 e-001 1,3785 e-001 1, 4442 e-001 1 , 5161 e-001 1.5934 e-001 4.6482 e + 001 4.6482 e + 001 4.6482 e + 001 4.6482 e + 001 4.6482 e + 001 4.6482 e + 001 4.6482 e + 001 4.6482 e + 001 4,6482 e + 001 4,6482 e + 001 4,6482 e + 001 4,6482 e + 001 4,6482 e + 001 4,6482 e + 001 4,6482 e + 001 4,6482 e + 001 4,6482 e + 001 4,6482 e + 001 4.6482 e + 001 2.6000 e-001 2.9000 e-001 3.2000 e-001 3.5000 e-001 3, 8000 e-001 4.10 O Oe-001 4.4000 e-001 4 , 7000 e-001, 00 00 e-001, 3000 e-001, 6000 e-001, 9000 e-001 6,2000 e-001 6,5000 e-001 6,8000 e-001 7,10 O Oe-001 7,4000 e-001 7,7000 e-001 8,00 00 e-001 7,1639 e-002 7,2801 e-002 7,3961 e-002 7,5246 e-002 7,6755 e-002 7,8571 e-002 8,0784 e-002 8 , 3490 e-002 8,6750 e-002 9,0536 e-002 9,4723 e-002 9,9123 e-002 1,0356 e-001 1,0791 e-001 1,1220 e-001 1,1657 e-001 1.2123 e-001 4.8500 e + 001 4, 8500 e + 001 4.8500 e + 001 4.8500 e + 001 4.8500 e + 001 4.8500 e + 001 4, 8500 e + 001 4.8500 e + 001 4 , 8500 e + 001 4.8500 e + 001 4.8500 e + 001 4, 8500 e + 001 4.8500 e + 001 4.8500 e + 001 4.8500 e + 001 4.8500 e + 001 4, 8500 e + 001 2.00 00 e- 001 2.3000 e-001 2.6000 e-001 2.9000 e-001 3.2000 e-001 3.5000 e-001 3.8000 e-001 4.10 O Oe-001 4.4000 e-001 4,7000 e-001, 00 00 e-001, 3000 e-001, 6000 e-001, 9000 e-001 6,2000 e-001 6,5000 e-001 6,8000 e-001 4,8500 e + 001 4,8500 e + 001 4,8500 e + 001 4,8500 e + 001 7, l OO Oe-001 7,4000 e-001 7,7000 e-001 8,0000 e-001 1,2643 e-001 1,3232 e -001 1.3894 e-001 1.4621 e-001

Table 2

  The first column (X) gives the minimum DNA size (Low) (in kilobases).

  The second column (Y) gives the switching time (in seconds).

  The third column (Z) gives the mobility of elec-

trophorèse. Data

X Y Z

48.5 30 0.3056

48.5 75 0.3150

48.5 90 0.3200

97.0 5 0.1540

97.0 10 0.2100

97.0 15 0.2719

97.0 30 0.2819

97.0 45 0.2790

97.0 70 0.2950

97.0 90 0.3067

97.0 105 0.3155

0.2256

145.5 30 0.2588

, 5, 45, 0.2605

, 5 60 0.2857

, 75, 0.2918

, 5 90 0.2942

145.5 105 0.3060

194.0 15 0.1677

194.0 30 0.2356

194.0 45 0.2437

194.0 60 0.2714

194.0 75 0.2767

194.0 90 0.2834

194.0 105 0.2960

.1157

0.1600

.2119

.2260

.2571

.2666

.2726

.2860

0.0925

.1863

.2084

.2452

0.2541

.2618

.2759

0.0690

.1528

.1907

.2310

.2440

.2510

.2660

0.1183

.1714

.2190

.2314

.2414

.2583

.0932

.1479

.2048

.2214

.2317

.2508

0.0789

0.1176

0.1905

242.5 242.5 242.5 242.5

242.5

242.5 242.5 242.5 291.0

291.0

291.0 291.0 291.0 291.0

291.0

339.5,

339.5 339.5 339.5

339.5

339.5 339.5 388.0 388.0

388.0

388.0 388.0 388.0 436.5

436.5

436.5 436.5 436.5 436.5

485.0

485.0 485.0

485.0 75 0.2098

485.0 90 0.2215

485.0 105 0.2408

533.5 40 0.0750

533.5 45 0.0924

533.5 60 0.1762

533.5 75 0.1987

533.5 90 0.2101

533.5 105 0.2333

582.0 40 0.0550

582.0 45 0.0748

582.0 60 0.1595

582.0 75 0.187

582.0 90 0.2011

582.0 105 0.2257

630.5 40 0.0400

* 630.5 45 0.0639

630.5 60 0.1333

630.5 75 0.1786

630.5 90 0.1931

630.5 105 0.2157

679.0 60 0.1095

679.0 75 0.1660

679.0 90 0.1863

679.0 105 0.2057

727.5 60 0.0881

727.5 75 0.1459

727.5 90 0.1766

776.0 60 0.0714

776.0 75 0.1233

776.0 90 0.1647

832.0 60 0.0620

832.0 75 0.0950

832.0 90 0.1425

832.0 105 0.1800

832.0 120 0.1837

915.0 75 0.0705

0.1070

.1650

.1782

  The first column (X) gives the minimum size

of DNA (Low) (kilobases).

  The second column (Y) gives the switching time

(in seconds)

The third

Electrophoresis.

Adapted surface

X Y

  column (Z) gives the mobility z 4,8500 e + 001 4,8500 e + 001 4,8500 e + 001 4,8500 e + 001 4,8500 e + 001 4,8500 e + 001 4,8500 e + 001 4,8500 e + 001 4, 8500 e + 001 4.8500 e + 001 4.8500 e + 001 4.8500 e + 001 4.8500 e + 001 4.8500 e + 001 4.8500 e + 001 4.8500 e + 001 4.8500 e + 001 4.8500 e + 001 4, 8500 e + 001 4.8500 e + 001 4.8500 e + 001, 0000 e + 000 1.0750 e + 001 1.6500 e + 001 2.2250 e + 001 2.8000 e + 001 3.3750 e + 001 3.9500 e + 001 4.5250 e + 001, 1000 e + 001, 6750 e + 001 6,2500 e + 001 6,8250 e + 001 7,4000 e + 001 7,9750 e + 001 8,5500 e + 001 9,1250 e + 001 9,7000 e + 001 1, 0275 e + 002 1,0850 e + 002 1,1425 e + 002 1,2000 e + 002 2,1572 e-001 2, 4990 e-001 2,7524 e-001 2,8992 e-001 3,0062 e-001 3,0761 e- 001 3,0930 e- 001 3,0986 e-001 3,1237 e-001 3,1459 e-001 3,1518 e-001 3,1519 e-001 3, 1607 e-001 3,1763 e-001 3 , 1890 e-001 3,1923 e-001 3,1973 e-001 3,2037 e-001 3,1918 e-001 3,1534 e-001 3,0926 e-001 915.0 915.0 915.0 9,1825 e + 001 9,1825 e + 001 9,1825 e + 001 9,1825 e + 001 9,1825 e + 001 9,1825 e + 001 9, 1825 e + 001 9,1825 e + 001 9,1825 e + 001 9,1825 e + 001 9,1825 e + 001 9,1825 e + 001 9,1825 e + 001 9,1825 e + 001 9,1825 e + 001 9,1825 e + 001 9, 1825 e + 001 9,1825 e + 001 9,1825 e + 001 9,1825 e + 001 9,1825 e + 001 1,3515 e + 002 1,3515 e + 001 1,3515 e + 001 1,3515 e + 001 1,3515 e + 002 1, 3515 e + 002 1.3515 e + 002 1.3515 e + 002 1.3515 e + 002 1.3515 e + 002 1.3515 e + 002 1.3515 e + 002 1.3515 e + 002 1.3515 e + 002 1.3515 e + 002, OOO Oe + 000 1,0750 e + 001 1,6500 e + 001 2,2250 e + 001 2,8000 e + 001 3,3750 e + 001 3,9500 e + 001 4,5250 e + 001, 100 l OO Oe + 001, 6750 e + 001 6,2500 e + 001 6,8250 e + 001 7,4000 e + 001 7,9750 e + 001 8,5500 e + 001 9,1250 e + 001 9,7000 e + 001 1,0275 e + 002 1,0850 e + 002 1,1425 e + 002 1,2000 e + 002, OOO Oe + 000 1,0750 e + 001 1,6500 e + 001 2,2250 e + 001 2,8000 e + 001 3,3750 e + 001 3,9500 e + 001 4,5250 e + 001, 100 l OO Oe + 001, 6750 e + 001 6,2500 e + 001 6,8250 e + 001 7,4000 e + 001 7,9750 e + 001 8,5500 e + 001 1,8147 e-001 2,2043 e-001 2,4968 e-001 2,6541 e-001 2, 7712 e-001 2.8517 e-001 2.8790 e-001 2.9026 e-001 2.9533 e-001 2.9984 e-001 3.0209 e-001 3.0310 e-001 3.0507 e-001 3.0742 e- 001 3,0899 e-001 3, 0997 e-001 3,1170 e-001 3,1344 e-001 3,1302 e-001 3,0986 e-001 3,0448 e-001 1,5246 e-001 1 , 9044 e-001 2,2009 e-001 2,3880 e-001 2,5327 e-001 2, 6321 e-001 2,6808 e-001 2,7284 e-001 2,7968 e-001 2,8586 e-001 2,8994 e-001 2,9233 e-001 2,9471 e-001 2,9725 e-001 2,9905 e-001 1,3515 e + 002 1, 3515 e + 001 1,3515 e + 002 1, 3515 e + 002 1.3515 e + 002 1.3515 e + 002 1, 7848 e + 002 1.7848 e + 002 1.7848 e + 002 1.7848 e + 002 1.7848 e + 002 1, 7848 e + 002 1.7848 e + 002 1, 7848 e + 002 1.7848 e + 002 1.7848 e + 002 1, 7848 e + 002 1.7848 e + 002 1.7848 e + 002 1.7848 e + 002 1.7848 e + 002 1, 7848 e + 002 1.7848 e + 002 1, 7848 e + 002 1.7848 e + 002 1.7848 e + 002 1, 7848 e + 002 2.2180 e + 002 2.2180 e + 002 2.2180 e + 002 2.2180 e + 002 2, 2180 e + 002 2.2180 e + 002 2, 2180 e + 002 2.2180 e + 002 9.1250 e + 001 9, 7000 e + 001 1.0275 e + 002 1.0850 e + 002 1.1425 e + 002 1.2000 e + 002, OOO Oe + 000 1, 0750 e + 001 1.6500 e + 001 2.2250 e + 001 2.8000 e + 001 3, 3750 e + 001 3.9500 e + 001 4.5250 e + 001, 100 l OO Oe + 001, 6750 e + 001 6, 2500 e + 001 6, 8250 e + 001 7,4000 e + 001 7,9750 e + 001 8,5500 e + 001 9, 1250 e + 001 9,7000 e + 001 1,0275 e + 002 1,0850 e + 002 1,1425 e + 002 1, 2000 e + 002, OOO Oe + 000 1,0750 e + 001 1,6500 e + 001 2,2250 e + 001 2, 8000 e + 001 3,3750 e + 001 3,9500 e + 001 4,5250 e + 001 3,0058 e-001 3, 0308 e-001 3 , 0533 e-001 3.0552 e-001 3.0318 e-001 2.987 e-001 1.2463 e-001 1.5894 e-001 1.8783 e-001 2.1054 e-001 2.2902 e-001 2,4163 e-001 2, 4907 e-001 2,5618 e-001 2,6458 e-001 2,7215 e-001 2,7777 e-001 2,8143 e-001 2,8420 e 001 2,8702 e-001 2,8921 e-001 2,9123 e-001 2,9419 e-001 2, 9677 e-001 2,9744 e-001 2,9582 e-001 2,9215 e-001 9 , 3584 e-002 1.2529 e-001 1.5495 e-001 1.8200 e-001 2.0457 e-001 2.2010 e-001 2.3015 e-001 2.3953 e-001 2.4955 e-001 2,5853 e-001 2,6543 e-001 2,7011 e-001 2,7355 e-001 2,7685 e-001 2,7947 e-001 2,8193 e-001 2,8525 e 001 2,8811 e-001 2, 8917 e -001 2,8816 e-001 2,8526 e-001 6,2018 e-002 9,3450 e-002 1,2447 e-001 1,5449 e-001 1,8017 e-001 1,9835 e-001 2,11 O Oe-001 2,2266 e-001 2, 3443 e-001 2,4496 e-001 2,5313 e-001 2,5872 e-001 2,6291 e-001 2,6678 e- 001 2 , 6980 e-001 2.7268 e-001 2.7636 e-001 2.7948 e-001 2.8090 e-001 2, 8045 e-001 2.7824 e-001, 1000 e + 001, 6750 e + 001 6 , 2500 e + 001 6,8250 e + 001 7,4000 e + 001 7,9750 e + 001 8,5500 e + 001 9,1250 e + 001 9,7000 e + 001 1,0275 e + 002 1,0850 e + 002 1,1425 e + 002 1 , 2000 e + 002, 0000 e + 000 1.0750 e + 001 1.6500 e + 001 2.2250 e + 001 2.8000 e + 001 3.3750 e + 001 3, 9500 e + 001 4.5250 e + 001, 1000 e + 001, 6750 e + 001 6,2500 e + 001 6,8250 e + 001 7,4000 e + 001 7,9750 e + 001 8,5500 e + 001 9,1250 e + 001 9,7000 e + 001 1,0275 e + 002 1,0850 e + 002 1,1425 e + 002 1,2,000 e + 002 2,2180 e + 002 2,2180 e + 002 2,2180 e + 002 2,2180 e + 002 2,2180 e + 002 2,2180 e + 002 2,2180 e + 002 2,2180 e + 002 2,2180 e + 002 2,2180 e + 002 2,2180 e + 002 2,2180 e + 002 2,2180 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 2,6512 e + 002 3,2160 e-002 3,0845 e + 002, 0000 e + 000 3.0845 e + 002 3.0845 e + 002 3.0845 e + 002 3.0845 e + 002 3.0845 e + 002 3.0845 e + 002 3.0845 e + 002 3.0845 e + 002 3.0845 e + 002 3.0845 e + 002 3.0845 e + 002 3.0845 e + 002 3.0845 e + 002 3.0845 e + 002 3, 0845 e + 002 3.0845 e + 002 3.0845 e + 002 3.0845 e + 002 3.0845 e + 002 3, 0845 e + 002 3.5178 e + 002 3.5178 e + 002 3.5178 e + 002 3.5178 e + 002 3, 5178 e + 002 3.5178 e + 002 3.5178 e + 002 3.5178 e + 002 3.5178 e + 002 3, 5178 e + 002 3.5178 e + 002 3.5178 e + 002 3.5178 e + 002 3.5178 e + 002 3, 5178 e + 002 3.5178 e + 002 1.0750 e + 001 1.6500 e + 001 2.2250 e + 001 2, 8000 e + 001 3.3750 e + 001 3.9500 e + 001 4.5250 e + 001, 1000 e + 001, 6750 e + 001 6.2500 e + 001 6.8250 e + 001 7.4000 e + 001 7.9750 e + 001 8 , 5500 e + 001 9,1250 e + 001 9,7000 e + 001 1,0275 e + 002 1,0850 e + 002 1,1425 e + 002 1,2000 e + 002, 0000 e + 000 1,0750 e + 001 1,6500 e + 001 2,2250 e + 001 2 , 8000 e + 001 3.3750 e + 001 3.9500 e + 001 4.5250 e + 001, 1000 e + 001, 6750 e + 001 6.2500 e + 001 6.8250 e + 001 7.4000 e + 001 7.9750 e + 001 8, 5500 e + 001 9,1250 e + 001 6,4833 e-002 9,6914 e-002 1,2816 e-001 1,5563 e-001 1,7604 e-001 1,9140 e-001 2,0546 e-001 2 , 1909 e-001 2.3130 e-001 2, 4079 e-001 2.479 e-001 2.5228 e-001 2.5676 e-001 2.6022 e-001 2.6352 e-001 2.6760 e-001 2.7103 e-001 2.7281 e-001 2.7288 e-001 2.7128 e-001 3, 8103 e-003 3.7852 e-002 7.0912 e-002 1.0275 e 001 1.3090 e-001 1.5313 e-001 1.7123 e-001 1.8771 e-001 2.0332 e-001 2.1738 e-001 2.2831 e-001 2, 3579 e-001 2 , 4 i 64 e-001 2.4680 e-001 2.5074 e-001 2.5451 e-001 2.5907 e-001 2.6291 e-001 2.6510 e-001 2.6562 e-001 2 , 6453 e-001 O, 0000 e + 000 1.2028 e-002 4.6202 e-002 7.8499 e-002 1.0673 e-001 1.3022 e-001 1.5045 e-001 1.6909 e -001 1,8689 e-001 2,0309 e-001 2,1562 e-001 2,2417 e -001 2, 3094 e-001 2.684 e-001 2.4135 e-001 2.4567 e-001 2.5075 e-001 2.5513 e-001 2.5778 e-001 2.5868 e-001 2,5799 e-001 O, 0000 e + 000 O, 0000 e + 000 2, 2665 e-002, 5437 e-002 8,3931 e-002 1,0788 e-001 1,2905 e-001 1,4937 e- 001 1.6958 e-001 3.5178 e + 002 3.5 178 e + 002 3.5178 e + 002 3.5178 e + 002 3.5178 e + 002 3.9510 e + 002 3.9510 e + 002 3.9510 e + 002 3, 9510 e + 002 3.9510 e + 002 3.9510 e + 002 3.9510 e + 002 3.9510 e + 002 3.9510 e + 002 3.9510 e + 002 3.9510 e + 002 3.9510 e + 002 3.9510 e + 002 3, 9510 e + 002 3.9510 e + 002 3.9510 e + 002 3.9510 e + 002 3.9510 e + 002 3.9510 e + 002 3.9510 e + 002 3.9510 e + 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e + 001 6,8250 e + 001 7,4000 e + 001 7, 9750 e + 001 8.5500 e + 001 9.1250 e + 001 9.700 e + 001 1.0275 e + 002 1.0850 e + 002 1.1425 e + 002 1.2000 e + 002 1.8823 e-001 2.0255 e-001 2,1237 e-001 2,2023 e-001 2,2689 e-001 2,3197 e-001 2,3687 e-001 2,4249 e-001 2,4741 e-001 2,5055 e-001 2, 5186 e-001 2.5156 e-001 O, 0000 e + 000 O, 0000 e + 000 O, 0000 e + 000 3.248 e-002 6.166 e-002 8.5536 e-002 1, 0691 e-001 1 , 2873 e-001 1.5156 e-001 1.7284 e-001 1.8908 e-001 2.0030 e- 001 2,0936 e-001 2,1687 e-001 2,2254 e-001 2,2801 e-001 2,3421 e-001 2, 3962 e-001 2,4323 e-001 2,4503 e-001 2 , 4515 e-001 4,3843 e + 002 4,3843 e + 002 4,3843 e + 002 4,3843 e + 002 4,3843 e + 002 4,3843 e + 002 4,3843 e + 002 4,3843 e + 002 4,3843 e + 002 4,3843 e + 002 4,3843 e + 002 4,3843 e + 002 4,8175 e + 002 4,8175 e + 002 4,8175 e + 002 4,8175 e + 002 4,8175 e + 002 4,8175 e + 002 4,8175 e + 002 4.8175 e + 002 4.8175 e + 002 4.8175 e + 002 4.8175 e + 002 4.8175 e + 002 4.8175 e + 002 4.8175 e + 002 4.8175 e 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+ 000 0, OOO Oe + 000 0.0000 e + 000 0, OOO Oe + 000 0.0000 e + 000 0, OOO Oe + 000 0, OOO Oe + 000 0, 0000 e + 000 O, 0000 e + 000 0, 0000 e + 00 0, 0 0000 e + 000 0, OOO O e + 000 O, 0000 e + 000 9,3618 e-003 3,2700 e-002, 4944 e-002 7,6071 e-002 9,5729 e-002 1,1341 e-001 1,2876 e-001 1,4168 e-001 0.0000 e + 000 O, 0000 e + 000 0, OOO Oe + 000 O, 0000 e + 000 0, OOO Oe + 000 0000 e + 000 0.0000 e + 000 0, OOO Oe + 000 1.1750 e + 003 1.1750 e + 003 1.1750 e + 003 1.1750 e + 003 1, 1750 e + 003 1.1750 e + 003 1.1750 e + 003 1,1750 e + 003 1,1750 e + 003 1, 1750 e + 003 1,1750 e + 003 1,1750 e + 003 1,1750 e + 003, 100 e + 001, 6750 e + 001 6,2500 e + 001 6,8250 e + 001 7, 4000 e + 001 7.9750 e + 001 8.5500 e + 001 9.1250 e + 001 9,7000 e + 001 1,0275 e + 002 1,0850 e + 002 1,1425 e + 002 1,2000 e + 002 O, 000 e + 00 O 0, OOO Oe + 00 OO, OO Oe + 00 O 0, OOO Oe + OOO 0, OOO Oe + 00 O 0, OOO Oe + OOO 1,8498 e-002 4,1753 e-002 6,3685 e-002 8,4069 e-002 1,0238 e-001 1,1896 e-001 1,3308 e -001

Table 3

  The first column (X) gives the DNA size (in kilobases).

  The second column (Y) gives the mobility (in centi-

meter per hour).

X Y

260 0.280

335 0.269

427 0.260

581 0.243

681 0.233

774 0.219

832 0.209

913 0.200

1051 0.173

1600 0.129

2200 0.100

2500 0.085

Claims (7)

  1 Process for improving the separation of macromolecules, in an electrophoresis system, using a computer, characterized in that it comprises the following steps: the capture of a first size representing the smallest macromolecule to solve, the seizure of a second size representing the largest macromolecule to solve,   memorization of a knowledge base   of calibration data obtained by separating macromolecules of known size under conditions that optimize separations, using the first size and second size for querying the knowledge base with creation of a set of optimized parameters, and   calibration of the system with the optimum parameters   wagered, so that the separation of macromolecules is optimized. Process according to Claim 1, characterized in   what the step of using sizes includes using   the first and second size for access to the knowledge base, such that an angle, a travel distance, a voltage gradient, an initial switch-on time, a final switch-on time and a   journey times are selected.   A method according to claim 2, characterized in that the calibration step comprises calibrating the system to provide optimum separation by use of angle, travel distance, voltage gradient, initial switching time, from   final switching time and travel time.   Method according to claim 1, characterized in that the step of storing a knowledge base comprises: storing a first set of relations   between the sizes of macromolecules and the times of   storage of a second set of relations between the sizes of the macromolecules and their mobility, and the memorization of a third set of relationships between minimum sizes of macromolecules, switching and mobility.   Electrophoresis system for improving the separation of macromolecules, characterized in that it comprises: an electrophoresis device (130), a computer (150) having a memory (107) and a processor (108), the computer being selectively coupled to the electrophoresis device,   an input device (102, 103) coupled to the computer   to enter a first size   of the smallest macromolecule that needs to be solved and the seizure of a second representative size of the largest molecule that needs to be solved, a knowledge base (120), coupled with the   memory and intended to retain information   Calibration data obtained by separating molecules of determined size under the conditions that optimize the separations, a programming device coupled to the memory and intended to indicate to the processor to use the first size and the second size for the access to the database   knowledge with creation of a set of optimal parameters wagered, and   a device for calibrating the electronic device   phoresis with optimized parameters so that the   separation of macromolecules is optimized.   System according to claim 5, characterized in   the programming device includes a   sitive coupled to the memory (107) for instructing the processor (108) to use the first size and second size for access to the knowledge base (120) and thereby to select an angle, a course distance , a voltage gradient, an initial switching time, a final switching time and a travel time. 7 System according to claim 6, characterized in that the calibration device comprises a device for calibrating the system so that it ensures optimum separation by use of the angle, the travel distance, the voltage gradient, the initial switching time, the final switching time and the travel time.   System according to claim 5, characterized in that   the knowledge base (120) includes a provision   storage device coupled to the memory and for maintaining a first set of relationships between macromolecule sizes and switching times, a   second set of relations between macromo-   lecules and mobilities, and a third set of relationships between minimum sizes of macromolecules,   switching times and mobilities.   9 System according to claim 5, characterized in that it further comprises a representation device for receiving the optimized parameters and coupled   selectively to the computer through an external interface.   System according to claim 7, characterized in   the electrophoresis device (130) is a device   electrophoresis system of homogeneous electric field type locked to the outline (CHEF).