CN210620780U - 1536 well solid phase synthesis and extraction plate - Google Patents
1536 well solid phase synthesis and extraction plate Download PDFInfo
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Abstract
The utility model discloses a 1536 hole solid phase synthesis and extraction board, be used for ultramicro DNA synthesis and compound solid phase extraction, including solid-phase carrier and cavity board, the cavity board has the cavity board body and locates 1536 filtration pores on this cavity board body, the filtration pore includes the square chamber portion that from the top down set gradually, transition chamber portion and circular chamber portion, square chamber portion upper portion is equipped with square entry, square chamber portion lower part is equipped with square body assembly position, square body assembly position is used for the square solid-phase carrier of holding, circular chamber portion upper portion is equipped with cylinder assembly position, cylinder assembly position is used for the circular solid-phase carrier of holding, circular chamber portion lower part is equipped with the export, and square chamber portion, the inner chamber sectional area of transition chamber portion and circular chamber portion diminishes gradually. The utility model provides a 1536 hole solid phase synthesis and extraction board have high flux, can handle characteristics such as super minim DNA synthesis and super minim compound solid phase extraction, still have leakproofness and compatible good, stable in quality, efficient, characteristics with low costs, therefore have wide market prospect.
Description
Technical Field
The utility model relates to the technical field of ultramicro DNA synthesis and compound solid phase extraction, in particular to a 1536-hole solid phase synthesis and extraction plate.
Background
Solid-Phase Extraction (SPE) is a sample pretreatment technology developed in recent years, is developed by combining Solid-liquid Extraction and column liquid chromatography, is mainly used for separating, purifying and concentrating samples, can improve the recovery rate of analytes compared with the traditional liquid-liquid Extraction method, more effectively separates the analytes from interfering components, reduces the sample pretreatment process, and is simple to operate, time-saving and labor-saving. The method is widely applied to the fields of medicine, food, environment, commodity inspection, chemical industry and the like.
At present, a DNA synthesis column and a solid phase extraction column usually adopt single-column tubes or 96-hole filter plates, the existing single-column tubes have multiple specifications and large volumes, are not beneficial to batch, large-scale and miniaturized operation, and are difficult to realize for micro, ultra-micro and large-scale DNA synthesis and compound solid phase extraction; the existing 96-hole filter plate mainly has two specifications of 1.5 and 2ml, can complete the synthesis work of 96 DNA samples, is more rapid than a single tube for the mass DNA synthesis and the solid-phase extraction of compounds, but cannot meet the requirements of mass and scale synthesis of micro and ultra-micro DNA and the solid-phase extraction of micro and ultra-micro compounds on the sample amount to be processed. The disadvantages of the current single column tube and 96-well filter plate are also shown in the following aspects: 1) the synthesis or solid phase extraction sample size is limited, and one experiment can synthesize 192 DNA samples at most and process 192 compound samples at the same time; 2) for the synthesis of DNA, the DNA synthesis scale of the existing product is more than 25nmol, and the synthesis of trace and ultra-trace DNA cannot be met, so that the dosage of the reagent for DNA synthesis required to be matched is very large, the reagent consumption is increased, the treatment cost is increased, most of the used treatment reagents are corrosive liquid which is toxic, harmful and volatile to human bodies and the environment, the more the dosage is, the more the waste liquid is discharged, and the more the harm to the human bodies and the environment is; 3) for the solid-phase extraction of compounds, the sample size of the treatment of the existing product is more than 1ml, and the solid-phase extraction of trace, ultra-trace or even trace compounds cannot be met, the existing product is adopted, so that not only is the reagent consumption increased, but also the treatment cost is increased, and most of the used treatment reagents are corrosive liquids which are toxic, harmful and volatile to human bodies and the environment, the more the dosage is, the more the waste liquid is discharged, and the greater the harm to the human bodies and the environment is; 4) the automation degree is not high, the efficiency is not high, the operation is very inconvenient, and the automatic and mass DNA synthesis and the solid-phase extraction of compound samples cannot be well realized; 5) the product has large volume, and more required matched reagents and filling matrixes are needed, so the production cost of the product is high; 6) the operation is complex, the function is single, time and labor are wasted, the cost and resources are wasted, and the requirement of integration of DNA synthesis and compound solid phase extraction cannot be met.
However, with the rapid development of biotechnology, life science, synthetic biology, biomedicine, chemical analysis, food detection and clinical diagnostics, a large amount of DNA samples need to be synthesized, a large amount of compound samples need to be subjected to solid phase extraction, and higher requirements are placed on related matched instruments and reagent consumables, such as high-throughput, ultra-small amount of DNA synthesis, compound solid phase extraction and detection, cost control, environmental protection and the like, and the usage amount of DNA synthesis, compound solid phase extraction reagents and the like is reduced to the maximum extent.
SUMMERY OF THE UTILITY MODEL
The utility model provides a 1536 hole solid phase synthesis and extraction board has solved the weak point that current single-column tube or 96 hole filter exist.
The technical scheme of the utility model is realized like this:
the embodiment of the utility model discloses 1536 hole solid phase synthesis and extraction board for ultramicro DNA synthesis and compound solid phase extraction, including solid phase carrier and cavity board, the cavity board has the cavity board body and locates 1536 filtration pores on this cavity board body, the filtration pore includes square chamber portion, transition chamber portion and the circular chamber portion that from the top down set gradually, square chamber portion upper portion is equipped with square entry, square chamber portion lower part is equipped with square body assembly position, square body assembly position is used for the square solid phase carrier of holding, circular chamber portion upper portion is equipped with cylinder assembly position, cylinder assembly position is used for the circular solid phase carrier of holding, circular chamber portion lower part is equipped with the export, just the inner chamber sectional area of square chamber portion, transition chamber portion and circular chamber portion diminishes gradually.
Further, the 1536 filter holes are arranged in a matrix of 48 rows of equally spaced filter holes in the transverse direction and 32 rows of equally spaced filter holes in the longitudinal direction.
Furthermore, the cylinder assembly position comprises a first cylinder assembly cavity and a second cylinder assembly cavity, the first cylinder assembly cavity is arranged at the upper end of the second cylinder assembly cavity, and the diameter of the section of the second cylinder assembly cavity is smaller than that of the section of the first cylinder assembly cavity.
Further, the transition cavity part is of a tapered structure with the gradually reduced inner cavity section along the direction from the square cavity part to the round cavity part.
Furthermore, the side length of the inner cavity of the cube assembling position is not more than 3.7mm, and the height of the inner cavity is 0.1-7 mm.
Furthermore, the diameter of the inner cavity of the cylinder assembly position is not more than 3.5mm, and the height of the inner cavity is 0.1 mm-7 mm.
Furthermore, the length range of the hollow plate is 120 mm-520 mm, the width range is 80 mm-360 mm, and the height range is 10 mm-50 mm.
Furthermore, the empty plate is provided with an identification block, and the identification block is used for displaying the placement direction of the empty plate.
Furthermore, an X-axis coding strip and a Y-axis coding strip are respectively arranged on one surface of the hollow plate close to the square inlet along the X-axis direction and the Y-axis direction.
Further, the outlet is of a bevel structure, a flat structure or a sharp structure.
The utility model provides a 1536 hole solid phase synthesis and extraction board has following beneficial effect:
1. the technical proposal of the utility model solves the difficult problem of the scale of synthesizing trace and ultra-trace DNA; the technical proposal of the utility model is specially designed for trace gene synthesis (0.01-10nmol), the synthesis scale is as low as 0.0001nmol, and the synthesis scale of the existing DNA synthesis column is generally 25nmol or more.
2. The technical proposal of the utility model solves the problem that the CPG carrier synthesized by DNA has overlarge volume; present CPG synthesizes the carrier, and diameter and thickness all are above 2mm basically, and the utility model discloses a 1536 hole solid phase synthesis and extraction board, solid phase synthesis carrier diameter and thickness are all within 2mm basically, even as low as 0.5 mm.
3. The technical proposal of the utility model solves the problem of excessive CPG usage in the solid phase synthesis carrier; the utility model discloses technical scheme is used for gene synthesis's carrier, and its CPG content just can satisfy gene synthesis's requirement between 1-50ug, however present solid phase synthesis carrier content is mostly more than 1mg, and some are more than 10mg even, and the DNA product of synthesizing out has far exceeded actual demand, has caused serious wasting of resources, has increased scientific research and manufacturing cost, is unfavorable for building saving type society, and the utility model discloses a 1536 hole solid phase synthesis and extraction board, CPG content can be as low as 0.01ug in its carrier.
4. The technical proposal of the utility model solves the problem of excessive reagent dosage in the DNA synthesis process, and saves the dosage of the synthesis reagent; the utility model discloses a 1536 hole solid phase synthesis and extraction board, solid phase synthesis carrier diameter and thickness are low to within 1.2mm, and used reagent volume is minimum to 0.1ul, and the required reagent volume of traditional synthetic carrier needs more than 5ul at least, and the synthetic reagent quantity of saving is more than 50 times at least. The technical scheme of the utility model not only reduced the reagent quantity, saved manufacturing cost, practiced thrift the resource, protected the environment.
5. The technical proposal of the utility model solves the difficult problem of solid phase extraction of trace and ultra-trace compounds; the technical scheme of the utility model can control the content of the function filler in 1536 hole solid phase synthesis and each hole in the extraction plate between 0.01-10mg, effectively reduced the use amount of function filler, and solid phase extraction carrier diameter and thickness all are basically within 2mm, even as low as 0.5mm for the sample volume of going up of compound solid phase extraction falls to 60ul and below, the use amount control of sample pretreatment reagent is at 10-60ul, save the reagent quantity more than 50 times than traditional solid phase extraction post. The technical scheme of the utility model not only reduced the reagent quantity, saved manufacturing cost, practiced thrift the resource, protected the environment.
6. The utility model discloses 1536 orifice plate leakproofness, compatible problem have been solved in technical scheme's design, make product quality more reliable.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without creative efforts.
Fig. 1 is a schematic view of an overall structure of a hollow plate according to an embodiment of the present invention;
fig. 2 is a schematic side view of a hollow plate according to an embodiment of the present invention;
FIG. 3 is a schematic cross-sectional view of a hollow plate according to an embodiment of the present invention;
fig. 4 is a schematic view of a structure of a middle filtering hole according to an embodiment of the present invention.
Detailed Description
The invention will be further understood by the following examples of embodiments of the invention, which are not intended to limit the invention. Insubstantial modifications and adaptations of the invention as described herein will occur to those skilled in the art and are intended to be within the scope of the invention.
The first embodiment is as follows:
1. 1536 solid phase synthesis and extraction plate for ultra-micro DNA synthesis and solid phase extraction of compounds, as shown in fig. 1, 2, 3 and 4, comprises a solid phase synthesis carrier (not shown) and a hollow plate 100, wherein the hollow plate 100 comprises a hollow plate body 1 and 1536 filter pores 2 arranged on the hollow plate body 1, 1536 filter pores 2 are arranged in a matrix of a row of 48 transverse equidistant filter pores and a row of 32 longitudinal equidistant filter pores, each filter pore 2 comprises a square cavity 21, a transition cavity 22 and a circular cavity 23 arranged in sequence from top to bottom, the upper part of the square cavity 21 is provided with a square inlet 211, the lower part of the square cavity 21 is provided with a square coordination 212, the square coordination 212 is used for accommodating the square solid phase synthesis carrier, the upper part of the circular cavity 23 is provided with a cylinder coordination, the cylinder coordination is used for accommodating the circular solid phase synthesis carrier, the lower part of the circular cavity 23 is provided with an outlet 24, and the square cavity 21, the square coordination 212, the transition cavity, The sectional areas of the inner cavities of the transition cavity part 22 and the circular cavity part 23 become smaller and smaller; wherein, the outline dimension of the blank plate 100 is: the length is 122.0mm, the width is 81.0mm, and the height is 42.0 mm; the width of the frame of the hollow plate 100 is 1.2mm, and the side length of the square cavity part 21 is gradually changed from 1.55mm of the side length of the square inlet 211 to 1.44mm of the side length of the cube assembly line 212; the transition cavity part 22 gradually closes up and gradually changes from a square hole with the side length of 1.44mm at the square body assembly position 212 to a circular hole until the diameter of the bottom part is 1.2 mm; the cylinder assembling position comprises a first cylinder assembling cavity 231 and a second cylinder assembling cavity 232, the first cylinder assembling cavity 231 is arranged at the upper end of the second cylinder assembling cavity 232, the diameter of the section of the second cylinder assembling cavity 232 is smaller than that of the section of the first cylinder assembling cavity 231, specifically, the first cylinder assembling cavity 231 is connected with the bottom of the transition cavity part 22, the diameter of the first cylinder assembling cavity 231 is 1.2mm, and the height of the first cylinder assembling cavity 231 is 2.5 mm; the diameter of the second cylinder assembly cavity 232 is 1.0mm, the height of the second cylinder assembly cavity is 1.2mm, and the joint between the first cylinder assembly cavity 231 and the second cylinder assembly cavity 232 is a horn-shaped transition area; the diameter of the outlet 24 at the lower end of the second cylinder assembly chamber 232 is 1.0 mm.
The blank 100 is provided with an identification block 3, the identification block 3 is used for displaying the placement direction of the blank 100, and one surface of the blank 100 close to the square inlet 211 is respectively provided with an X-axis coding strip 4 (horizontal coordinates are marked by Arabic numerals) and a Y-axis coding strip 5 (vertical coordinates are marked by English letters) along the X-axis direction and the Y-axis direction.
2. Preparation of solid-phase synthetic support (10000 solid-phase synthetic supports per preparation):
1) selecting raw materials: the CPG (i.e. microporous Glass bead Controlled Pore Glass, abbreviated as CPG, hereinafter referred to as the same meaning) loading capacity is 20umol/g, and the CPG content of each 0.1nmol solid phase synthesis carrier is about 5 ug; the molecular weight of the UHMW-PE powder (which is ultra-high molecular weight polyethylene and is synonymous hereinafter) is 300 ten thousand.
2) Calculating the volume of the raw materials: the volume of the synthesized carrier with each side of 1.44mm and the thickness of 1.0mm is 2.0736ul, the synthesized carrier is prepared according to 10000 particles, and the total volume is 20.736ml and approximately equals to 20.7 ml.
3) Preparing a sintering raw material of the synthetic carrier: weighing 50mg CPG, putting into a 50ml measuring cylinder, slowly adding UHMW-PE powder to 20.7ml scale, reversing and uniformly mixing for several times, and then filling up to 20.7ml position with UHMW-PE; repeatedly reversing and mixing evenly for later use.
4) Installing a mold: uniformly adding mixed CPG and UHMW-PE blended powder into an aluminum mold containing 10000 holes with side length of 1.44mm and height of 1.0mm, vibrating the mixture by using a vibrator, covering an upper cover plate, and locking.
5) And (3) sintering: sintering at 200 deg.C for 35min, cooling with water, and taking out solid phase synthesis carrier.
3. Assembly of solid phase synthesis supports
The method comprises the steps of assembling a square solid-phase synthesis carrier with the side length of 1.44mm and the thickness of 1.0mm to a square assembly position with the side length of 1.44mm and the height of 1.2mm by adopting full-automatic filling equipment.
The 1536-well solid phase synthesis and extraction plate of this example was used as a 0.1nmol 1536-well DNA synthesis purification plate.
Example II:
1. 1536 solid phase synthesis and extraction plate for ultramicro DNA synthesis and compound solid phase extraction, the structure of the hollow plate is mostly the same as that of the hollow plate in the embodiment, the hollow plate comprises a solid-phase synthesis carrier and a hollow plate, the hollow plate is provided with a hollow plate body and 1536 filter holes arranged on the hollow plate body, 1536 filter holes are arranged in a row of 48 transverse equidistant filter holes and a row of 32 longitudinal equidistant filter holes in a matrix manner, each filter hole comprises a square cavity part, a transition cavity part and a circular cavity part which are sequentially arranged from top to bottom, a square inlet is arranged at the upper part of the square cavity part, a square assembly part is arranged at the lower part of the square cavity part and is used for accommodating the square solid-phase synthesis carrier, a cylinder assembly part is arranged at the upper part of the circular cavity part and is used for accommodating the circular solid-phase synthesis carrier, an outlet is arranged at the lower part of the circular cavity part, and the sectional areas of the inner cavities of the square cavity part; wherein, blank overall dimension: the length is 122.0mm, the width is 81.0mm, and the height is 42.0 mm; the side length of the square cavity part is 1.55mm from the square inlet, and gradually changes from the close to the square body assembling position to be 1.44 mm; the transition cavity part gradually closes and gradually changes from a square hole with the side length of 1.44mm at the square assembly position to a circular hole until the diameter of the bottom part is 1.2 mm; the cylinder assembly position comprises a first cylinder assembly cavity and a second cylinder assembly cavity, the first cylinder assembly cavity is arranged at the upper end of the second cylinder assembly cavity, the section diameter of the second cylinder assembly cavity is smaller than that of the first cylinder assembly cavity, specifically, the first cylinder assembly cavity is connected with the bottom of the transition cavity part, the diameter is 1.2mm, and the height is 2.5 mm; the diameter of the second cylinder assembly cavity is 1.0mm, the height of the second cylinder assembly cavity is 1.2mm, and the joint between the first cylinder assembly cavity and the second cylinder assembly cavity is a horn-shaped transition area; the outlet diameter at the lower end of the second cylinder assembly chamber is 1.0 mm.
2. Preparation of solid-phase synthetic support (10000 solid-phase synthetic supports per preparation):
1) raw materials: the CPG loading capacity is 40umol/g, and the CPG content of each solid-phase synthesis carrier of 5nmol is about 125 ug; the molecular weight of the UHMW-PE powder is 300 ten thousand.
2) Calculating the volume of the raw materials: the volume of each solid phase synthesis carrier with the diameter of 1.0mm and the thickness of 1.2mm is 0.942ul, the mixture ratio of 10000 particles is adopted, and the total volume is 9.42 ml.
3) Preparing a sintering raw material of a solid-phase synthesis carrier: weighing 1250mg of CPG, putting the CPG into a 10ml measuring cylinder, slowly adding UHMW-PE powder to 9.42ml scale, reversing and uniformly mixing for several times, and then filling the UHMW-PE powder to the 9.42ml position; repeatedly reversing and mixing evenly for later use.
4) Installing a mold: uniformly adding mixed CPG and UHMW-PE blended powder into an aluminum mould containing 10000 holes with the diameter of 1.0mm and the height of 1.2mm, vibrating the mixed powder by using a vibrator, covering an upper cover plate, and locking.
5) And (3) sintering: sintering for 28min at 200 ℃, cooling by water, and taking out the solid-phase synthesis carrier.
3. Assembly
And (3) assembling the solid-phase synthesis carrier with the diameter of 1.0mm and the thickness of 1.2mm to a second cylinder assembly cavity with the diameter of 1.0mm and the height of 1.2mm by adopting full-automatic filling equipment.
The 1536-well solid phase synthesis and extraction plate of this example was used as a 5nmol 1536-well DNA synthesis purification plate.
Example III:
1. 1536 solid phase synthesis and extraction plate for ultramicro DNA synthesis and compound solid phase extraction, the structure of the hollow plate is mostly the same as that of the hollow plate in the embodiment, the hollow plate comprises a solid-phase synthesis carrier and a hollow plate, the hollow plate is provided with a hollow plate body and 1536 filter holes arranged on the hollow plate body, 1536 filter holes are arranged in a row of 48 transverse equidistant filter holes and a row of 32 longitudinal equidistant filter holes in a matrix manner, each filter hole comprises a square cavity part, a transition cavity part and a circular cavity part which are sequentially arranged from top to bottom, a square inlet is arranged at the upper part of the square cavity part, a square assembly part is arranged at the lower part of the square cavity part and is used for accommodating the square solid-phase synthesis carrier, a cylinder assembly part is arranged at the upper part of the circular cavity part and is used for accommodating the circular solid-phase synthesis carrier, an outlet is arranged at the lower part of the circular cavity part, and the sectional areas of the inner cavities of the square cavity part; wherein, blank overall dimension: the length is 122.0mm, the width is 81.0mm, and the height is 42.0 mm; the side length of the square cavity part is 1.55mm from the square inlet, and gradually changes from the close to the square body assembling position to be 1.44 mm; the transition cavity part gradually closes and gradually changes from a square hole with the side length of 1.44mm at the square assembly position to a circular hole until the diameter of the bottom part is 1.2 mm; the cylinder assembly position comprises a first cylinder assembly cavity and a second cylinder assembly cavity, the first cylinder assembly cavity is arranged at the upper end of the second cylinder assembly cavity, the section diameter of the second cylinder assembly cavity is smaller than that of the first cylinder assembly cavity, specifically, the first cylinder assembly cavity is connected with the bottom of the transition part, the diameter is 1.2mm, and the height is 2.5 mm; the diameter of the second cylinder assembly cavity is 1.0mm, the height of the second cylinder assembly cavity is 1.2mm, and the joint between the first cylinder assembly cavity and the second cylinder assembly cavity is a horn-shaped transition area; the outlet diameter at the lower end of the second cylinder assembly chamber is 0.5 mm.
2. Preparation of solid phase Synthesis support (quantity of solid phase Synthesis support 10000 per preparation)
1) Raw materials: the CPG loading capacity is 20umol/g, and the CPG content of each solid phase synthesis carrier of 10nmol is about 500 ug; the molecular weight of the UHMW-PE powder is 300 ten thousand.
2) Calculating the volume of the raw materials: the volume of each solid phase synthesis carrier with the diameter of 1.2mm and the thickness of 2.5mm is 2.826ul, the mixture ratio is 10000, and the total volume is 28.26 ml.
3) Preparing a sintering raw material of a solid-phase synthesis carrier: weighing 5g of CPG, putting the CPG into a 50ml measuring cylinder, slowly adding UHMW-PE powder to 28.26ml of scale, reversing and uniformly mixing for several times, and then filling the UHMW-PE powder to 28.26 ml; repeatedly reversing and mixing evenly for later use.
4) Installing a mold: uniformly adding mixed CPG and UHMW-PE blended powder into an aluminum mold containing 10000 holes with the diameter of 1.2mm and the height of 2.5mm, vibrating the mixture by using a vibrator, covering an upper cover plate, and locking.
5) And (3) sintering: sintering for 45min at 200 ℃, cooling by water, and taking out the solid-phase synthesis carrier.
3. Assembly
And (3) assembling the solid-phase synthesis carrier with the diameter of 1.2mm and the thickness of 2.5mm to a first cylinder assembly cavity with the diameter of 1.2mm and the height of 2.5mm in the 1536-hole synthesis hollow plate by adopting full-automatic filling equipment.
The 1536-well solid phase synthesis and extraction plate of this example was used as a 10nmol 1536-well DNA synthesis purification plate.
Example four:
1. 1536 solid phase synthesis and extraction plate for ultramicro DNA synthesis and compound solid phase extraction, the structure of the hollow plate is mostly the same as that of the hollow plate in the embodiment, the hollow plate comprises a solid-phase synthesis carrier and a hollow plate, the hollow plate is provided with a hollow plate body and 1536 filter holes arranged on the hollow plate body, 1536 filter holes are arranged in a row of 48 transverse equidistant filter holes and a row of 32 longitudinal equidistant filter holes in a matrix manner, each filter hole comprises a square cavity part, a transition cavity part and a circular cavity part which are sequentially arranged from top to bottom, a square inlet is arranged at the upper part of the square cavity part, a square assembly part is arranged at the lower part of the square cavity part and is used for accommodating the square solid-phase synthesis carrier, a cylinder assembly part is arranged at the upper part of the circular cavity part and is used for accommodating the circular solid-phase synthesis carrier, an outlet is arranged at the lower part of the circular cavity part, and the sectional areas of the inner cavities of the square cavity part; wherein, blank overall dimension: 488.0mm in length, 324.0mm in width and 17.22mm in height; the side length of the square cavity part is 3.59mm from the square inlet, and gradually changes from the close to the square body assembling position to 3.5 mm; the transition cavity part gradually closes and gradually changes from a square hole with the side length of 3.5mm at the square assembly position to a circular hole until the diameter of the bottom part is 1.2 mm; the cylindrical assembly cavity is connected with the bottom of the transition cavity part, the diameter of the cylindrical assembly cavity is 1.2mm, and the height of the cylindrical assembly cavity is 1.5 mm; the lower end of the cylindrical assembly cavity is closed to form a tube body with the diameter of 0.8mm and the height of 14 mm.
2. Preparation of solid-phase Synthesis support (number of solid-phase Synthesis supports 5000 per preparation)
1) Raw materials: the CPG loading capacity is 25umol/g, and the CPG content of each 1nmol solid phase synthesis carrier is about 40 ug; the molecular weight of the UHMW-PE powder is 300 ten thousand.
2) Calculating the volume of the raw materials: the volume of each solid phase synthesis carrier with the diameter of 1.2mm and the thickness of 1.0mm is 1.1304ul, the total volume is 5.625ml and approximately equals to 5.6ml according to the proportion of 5000 particles.
3) Preparing a sintering raw material of a solid-phase synthesis carrier: weighing 200mg CPG, putting into a 10ml graduated cylinder, slowly adding UHMW-PE powder to the scale of 5.6ml, reversing and uniformly mixing for several times, and then filling the UHMW-PE powder to the position of 5.6 ml; repeatedly reversing and mixing evenly for later use.
4) Installing a mold: uniformly adding the mixed CPG and UHMW-PE blended powder into an aluminum mold containing 5000 holes with the diameter of 1.2mm and the height of 1.0mm, vibrating the mixture by using a vibrator, covering an upper cover plate, and locking.
5) And (3) sintering: sintering at 190 deg.C for 30min, cooling with water, and taking out solid phase synthesis carrier.
3. Assembly
And assembling the solid-phase synthesis carrier with the diameter of 1.2mm and the thickness of 1.0mm to a 1.2mm assembling position in the 1536-hole synthesis empty plate by adopting full-automatic filling equipment.
The 1536-well solid phase synthesis and extraction plate of this example was used as a 1nmol 1536-well DNA synthesis purification plate.
Example five:
1. 1536 solid phase synthesis and extraction plate for ultramicro DNA synthesis and compound solid phase extraction, the structure of the hollow plate is mostly the same as that of the hollow plate in the embodiment, the hollow plate comprises a solid-phase synthesis carrier and a hollow plate, the hollow plate is provided with a hollow plate body and 1536 filter holes arranged on the hollow plate body, 1536 filter holes are arranged in a row of 48 transverse equidistant filter holes and a row of 32 longitudinal equidistant filter holes in a matrix manner, each filter hole comprises a square cavity part, a transition cavity part and a circular cavity part which are sequentially arranged from top to bottom, a square inlet is arranged at the upper part of the square cavity part, a square assembly part is arranged at the lower part of the square cavity part and is used for accommodating the square solid-phase synthesis carrier, a cylinder assembly part is arranged at the upper part of the circular cavity part and is used for accommodating the circular solid-phase synthesis carrier, an outlet is arranged at the lower part of the circular cavity part, and the sectional areas of the inner cavities of the square cavity part; wherein, blank overall dimension: 488.0mm in length, 324.0mm in width and 17.22mm in height; the side length of the square cavity part is 3.59mm from the square inlet, and gradually changes from the close to the square body assembling position to 3.5 mm; the transition part gradually narrows from a square hole with the side length of 3.5mm to a circular hole until the diameter of the bottom part is 1.2 mm; the cylindrical assembly cavity is connected with the bottom of the transition cavity part, the diameter of the cylindrical assembly cavity is 1.2mm, and the height of the cylindrical assembly cavity is 1.5 mm; the lower end of the cylindrical assembly cavity is closed to form a tube body with the diameter of 0.5mm and the height of 14 mm.
2. Preparation of solid phase Synthesis support (calculated as 5000 particles prepared)
1) Raw materials: the CPG loading capacity is 60umol/g, and the CPG content of each 3nmol solid phase synthesis carrier is about 50 ug; the molecular weight of the UHMW-PE powder is 300 ten thousand.
2) Calculating the volume of the raw materials: the volume of each solid phase synthesis carrier with the diameter of 1.2mm and the thickness of 1.0mm is 1.1304ul, the total volume is 5.625ml and approximately equals to 5.6ml according to the proportion of 5000 particles.
3) Preparing a sintering raw material of a solid-phase synthesis carrier: weighing 250mg of CPG, putting the CPG into a 10ml measuring cylinder, slowly adding UHMW-PE powder to a scale of 5.6ml, reversing and uniformly mixing for several times, and then filling the UHMW-PE powder to a position of 5.6 ml; repeatedly reversing and mixing evenly for later use.
4) Installing a mold: uniformly adding the mixed CPG and UHMW-PE blended powder into an aluminum mold containing 5000 holes with the diameter of 1.2mm and the height of 1.0mm, vibrating the mixture by using a vibrator, covering an upper cover plate, and locking.
5) And (3) sintering: sintering at 190 deg.C for 35min, cooling with water, and taking out solid phase synthesis carrier.
3. Assembly
And assembling the solid-phase synthesis carrier with the diameter of 1.2mm and the thickness of 1.0mm to a 1.2mm assembling position in the 1536-hole synthesis empty plate by adopting full-automatic filling equipment.
The 1536-well solid phase synthesis and extraction plate of this example was used as a 3nmol 1536-well DNA synthesis purification plate.
Example six:
the 1536-hole C18 solid phase extraction plate has C18 powder with 0.3 mg/hole of functional filler content, and is divided into a cylindrical solid phase extraction carrier with the diameter of 1.2mm and the thickness of 1.0mm and a 1536-hole filter plate matched with the same.
1. Design of hollow plate with 1536-hole filter plate
1536 hole C18 solid phase extraction board for ultra-micro compound solid phase extraction, the structure of which is mostly the same as the structure of the hollow board in the first embodiment, including solid phase extraction carrier and hollow board, the hollow board has hollow board body and 1536 filter holes on the hollow board body, 1536 filter holes are arranged in a matrix by a row of 48 filter holes with equal distance transversely and a row of 32 filter holes with equal distance longitudinally, the filter holes include a square cavity part, a transition cavity part and a round cavity part which are arranged from top to bottom in sequence, the upper part of the square cavity part is provided with a square inlet, the lower part of the square cavity part is provided with a square assembly coordinate, the square assembly position is used for accommodating the square solid phase extraction carrier, the upper part of the round cavity part is provided with a cylinder assembly coordinate, the cylinder assembly position is used for accommodating the round solid phase extraction carrier, the lower part of the round cavity part is provided with an outlet, and the cross-sectional areas of the square cavity part; wherein, blank overall dimension: the length is 127.3mm, the width is 85.0mm, and the height is 26.0 mm; the side length of the square cavity part is 1.55mm from the square inlet, and gradually changes from the close to the square body assembling position to be 1.44 mm; the transition cavity part gradually closes and gradually changes from a square hole with the side length of 1.44mm at the square assembly position to a circular hole until the diameter of the bottom part is 1.2 mm; the cylinder assembly position comprises a first cylinder assembly cavity and a second cylinder assembly cavity, the first cylinder assembly cavity is arranged at the upper end of the second cylinder assembly cavity, the section diameter of the second cylinder assembly cavity is smaller than that of the first cylinder assembly cavity, specifically, the first cylinder assembly cavity is connected with the bottom of the transition part, the diameter is 1.2mm, and the height is 1.0 mm; the diameter of the second cylinder assembly cavity is 1.0mm, the height of the second cylinder assembly cavity is 1.2mm, and the joint between the first cylinder assembly cavity and the second cylinder assembly cavity is a horn-shaped transition area; the outlet diameter at the lower end of the second cylinder assembly chamber is 0.5 mm.
2. Preparation of solid phase extraction Carriers (calculated by the number of 5000 solid phase extraction Carriers prepared)
1) Raw materials: calculated as 0.3mg of C18 powder functional filler per hole, 5000 required C18 powder functional fillers were 1.5g, and the molecular weight of the UHMW-PE powder was 300 ten thousand.
2) Calculating the volume of the raw materials: the volume of each solid phase synthesis carrier with the diameter of 1.2mm and the thickness of 1.0mm is 1.1304ul, the total volume is 5.625ml and approximately equals to 5.6ml according to the proportion of 5000 particles.
3) Preparing a solid phase extraction carrier sintering raw material: weighing 1.5g C18 powder functional filler, placing into a 10ml graduated cylinder, slowly adding UHMW-PE powder to 5.6ml scale, reversing and mixing uniformly for several times, and then filling up to 5.6ml position with UHMW-PE; repeatedly reversing and mixing evenly for later use.
4) Installing a mold: uniformly adding the mixed C18 and UHMW-PE blended powder into an aluminum mold containing 5000 holes with the diameter of 1.2mm and the height of 1.0mm, vibrating the powder by a vibrator to be flat, covering an upper cover plate, and locking.
5) And (3) sintering: sintering at 190 deg.C for 35min, water cooling, and taking out C18 solid phase extraction carrier.
3. Assembly
The solid phase extraction carrier with the diameter of 1.2mm and the thickness of 1.0mm is assembled to the assembly position of 1.2mm in the 1536-hole filtering empty plate by adopting full-automatic filling equipment, namely the plate can be used as a 1536-hole C18 solid phase extraction plate.
Example seven:
1536-well nucleic acid extraction plate contains Silica 0.25 mg/well and DNA 0-5ug, and is divided into cylindrical Silica gel solid phase carrier with diameter of 1.2mm and thickness of 2.5mm and 1536-well filter plate.
1. Design of hollow plate with 1536-hole filter plate
1536 hole nucleic acid extraction board for DNA extraction of ultramicro nucleic acid sample, the structure of which is mostly the same as that of the hollow board in the first embodiment, comprises solid phase nucleic acid extraction carrier and hollow board, the hollow board has hollow board body and 1536 filter holes arranged on the hollow board body, 1536 filter holes are arranged in a matrix by a row of 48 filter holes with equal distance transversely and a row of 32 filter holes with equal distance longitudinally, the filter holes comprise a square cavity part, a transition cavity part and a round cavity part which are arranged from top to bottom in sequence, the upper part of the square cavity part is provided with a square inlet, the lower part of the square cavity part is provided with a square assembly coordinate, the square assembly position is used for accommodating the square solid phase nucleic acid extraction carrier, the upper part of the round cavity part is provided with a cylinder assembly coordinate, the cylinder assembly position is used for accommodating the round solid phase nucleic acid extraction carrier, the lower part of the round cavity part is, the sectional areas of the inner cavities of the transition cavity part and the circular cavity part are gradually reduced; wherein, blank overall dimension: the length is 127.3mm, the width is 85.0mm, and the height is 26.0 mm; the side length of the square cavity part is 1.55mm from the square inlet, and gradually changes from the close to the square body assembling position to be 1.44 mm; the transition cavity part gradually closes and gradually changes from a square hole with the side length of 1.44mm at the square assembly position to a circular hole until the diameter of the bottom part is 1.2 mm; the cylinder assembly position comprises a first cylinder assembly cavity and a second cylinder assembly cavity, the first cylinder assembly cavity is arranged at the upper end of the second cylinder assembly cavity, the section diameter of the second cylinder assembly cavity is smaller than that of the first cylinder assembly cavity, specifically, the first cylinder assembly cavity is connected with the bottom of the transition part, the diameter is 1.2mm, and the height is 2.5 mm; the diameter of the second cylinder assembly cavity is 1.0mm, the height of the second cylinder assembly cavity is 1.2mm, and the joint between the first cylinder assembly cavity and the second cylinder assembly cavity is a horn-shaped transition area; the outlet diameter at the lower end of the second cylinder assembly chamber is 0.5 mm.
2. Preparation of solid Supports (calculated on the number of 5000 solid Supports prepared)
1) Raw materials: 5000 particles of the desired Silica filler were 1.25g calculated as 0.25mg Silica per pore and the molecular weight of the UHMW-PE powder was 400 ten thousand.
2) Calculating the volume of the raw materials: the volume of each solid phase carrier with the diameter of 1.2mm and the thickness of 2.5mm is 2.826ul, the proportion of each solid phase carrier is 5000 particles, and the total volume is 14.13 ml.
3) Preparing solid phase carrier sintering raw materials: weighing 1.25g of silicon dioxide Silica filler, putting the silicon dioxide Silica filler into a 20ml measuring cylinder, slowly adding UHMW-PE powder to a scale of 14.13ml, reversing and uniformly mixing for several times, and then filling up the UHMW-PE powder to a position of 14.13 ml; repeatedly reversing and mixing evenly for later use.
4) Installing a mold: uniformly adding mixed silicon dioxide Silica filler and UHMW-PE blended powder into an aluminum mold containing 5000 holes with the diameter of 1.2mm and the height of 2.5mm, vibrating the mixture by using a vibrator, covering an upper cover plate, and locking.
5) And (3) sintering: sintering at 190 deg.C for 35min, water cooling, and taking out solid phase carrier.
3. Assembly
The solid phase carrier with the diameter of 1.2mm and the thickness of 2.5mm is assembled to a 1.2mm assembly position in a 1536-hole filtering empty plate by adopting full-automatic filling equipment.
4. Coupling of hydroxyl functional chemical groups:
placing the 1536-hole filter plate filled with the solid phase carrier into a sealing hole matched with a negative pressure suction filtration device, and gradually adding 40ul of concentrated sulfuric acid in volume ratio into the 1536-hole filter plate filled with the solid phase carrier by adopting a piranha method for three times under the coordination of a sample adding and separating manipulator: H2O2 ═ 2: 1, reacting at 85 ℃ for 0.5h, activating hydroxyl groups needed by people in unbonded Silica gel, cleaning, heating to 80 ℃, preserving heat for 5 hours, and drying, wherein the Silica powder coupling hydroxyl becomes Silica functional filler, and can be used as a nucleic acid extraction or solid phase extraction column plate after activation, and the structure is as follows:
(Note: piranha solution is a hot mixture of concentrated sulfuric and hydrogen peroxide to remove all organics, while hydroxylating most of the material surface. H2SO4+ H2O2 → H3O + + HSO4- + O atomic oxygen is a very strong oxidant.)
That is, the plate can be used as a 1536-well nucleic acid extraction plate.
Example eight:
the 1536-hole WCX (cation exchange) solid phase extraction plate has the polystyrene PS powder filler content of 0.2 mg/hole, and is divided into a square solid phase carrier with the side length of 1.44mm and the thickness of 1.2mm and a 1536-hole filter plate matched with the square solid phase carrier.
1. Design of hollow plate with 1536-hole filter plate
1. 1536 hole solid phase synthesis and extraction board for micro DNA synthesis and compound solid phase extraction, the structure is mostly the same as the structure of a hollow board of the embodiment, including solid phase carrier and hollow board, the hollow board has a hollow board body and 1536 filter holes arranged on the hollow board body, 1536 filter holes are arranged in a row according to 48 transverse equidistant filter holes and a row of 32 longitudinal equidistant filter holes in a matrix, the filter holes include a square cavity part, a transition cavity part and a round cavity part which are arranged from top to bottom in sequence, the upper part of the square cavity part is provided with a square inlet, the lower part of the square cavity part is provided with a square assembly coordinate, the square assembly position is used for accommodating the square carrier, the upper part of the round cavity part is provided with a cylinder assembly coordinate, the cylinder assembly position is used for accommodating the round solid phase carrier, the lower part of the round cavity part is provided with an outlet, and the cross-sectional areas of the square cavity part, the transition cavity part and the round.
2. Preparation of solid Supports (calculated on the number of 5000 solid Supports prepared)
1) Raw materials: calculated according to the polystyrene PS powder filler content of 0.2mg in each hole, 5000 required polystyrene PS powder fillers are 1g, and the molecular weight of the UHMW-PE powder is 400 ten thousand.
2) Calculating the volume of the raw materials: the volume of the solid phase carrier with each side of 1.44mm and the thickness of 1.2mm is 2.48832ul, the total volume is 12.4416ml and is approximately equal to 12.44ml according to the proportion of 5000 particles.
3) Preparing solid phase carrier sintering raw materials: weighing 1g of polystyrene PS powder filler, putting the polystyrene PS powder filler into a 20ml measuring cylinder, slowly adding UHMW-PE powder to 12.44ml of scale, reversing and uniformly mixing for several times, and then filling up to 12.44ml with UHMW-PE; repeatedly reversing and mixing evenly for later use.
4) Installing a mold: uniformly adding mixed polystyrene PS powder filler and UHMW-PE blended powder into an aluminum die containing 5000 holes with the side length of 1.44mm and the height of 1.2mm, vibrating the mixed powder by using a vibrator, covering an upper cover plate, and locking.
5) And (3) sintering: sintering at 190 deg.C for 35min, water cooling, and taking out solid phase carrier.
3. Assembly
The solid phase carrier with the side length of 1.44mm and the thickness of 1.2mm is assembled to a 1.44mm assembly position in a 1536-hole filtering empty plate by adopting full-automatic filling equipment.
4. Coupling of carboxyl functional chemical groups:
1) acetylation of Polystyrene (PS) cured sieve plate:
placing a 1536-hole filter plate filled with a solid phase carrier into a sealing hole matched with a negative pressure suction filtration device, sequentially adding 8ul of Dichloromethane (DCM) and 8ul of acetylation reagent (Reg) into the 1536-hole filter plate filled with the solid phase carrier under the coordination of a sample adding and separating manipulator, adding 15ul of catalyst (Cat) under the condition of reversely introducing nitrogen, reacting for 20min at normal temperature and normal pressure, after the reaction is finished, sequentially cleaning and suction-filtering by THF, glacial hydrochloric acid (3 percent) and distilled water until filtrate is free of chloride ions detected by a silver nitrate solution, then filtering and washing by methanol for 5 times, and drying in vacuum for later use.
2) Carboxylation of acetylated polystyrene PS cured sieve plate:
placing a 1536-hole filter plate filled with acetylated polystyrene PS solid phase carriers into a sealing hole matched with a negative pressure suction filtration device, adding 15ul of Dichloromethane (DCM) into the 1536-hole filter plate filled with the solid phase carriers under the coordination of a sample adding and separating manipulator, keeping the flow rate at 0.2d/min, repeatedly operating the steps for more than three times under the condition of ensuring the cleanness and the smoothness of a cylinder, and preparing bromine water, KOH and water into a bromine water alkali solution at 0 ℃ after the reaction is finished; adding the solution into a column tube, keeping the flow rate at 0.2d/min, repeatedly operating for more than three times under the condition of ensuring the cleanness and the smoothness of the column body, after the reaction is finished, respectively and fully washing, filtering and vacuum-drying by using diluted hydrochloric acid, distilled water, ethanol and methanol with proper volumes.
The reaction equation is shown in the following chart:
the plate was used as a 1536-well WCX (cation exchange) solid phase extraction plate.
The above description is only a preferred embodiment of the present invention, and should not be taken as limiting the invention, and any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A1536-hole solid phase synthesis and extraction plate is used for ultramicro DNA synthesis and compound solid phase extraction and is characterized by comprising a solid phase carrier and a hollow plate, wherein the hollow plate is provided with a hollow plate body and 1536 filter holes arranged on the hollow plate body, each filter hole comprises a square cavity part, a transition cavity part and a circular cavity part which are sequentially arranged from top to bottom, a square inlet is formed in the upper part of the square cavity part, a square assembling part is arranged at the lower part of the square cavity part and used for accommodating the square solid phase carrier, a cylinder assembling part is arranged at the upper part of the circular cavity part and used for accommodating the circular solid phase carrier, an outlet is formed in the lower part of the circular cavity part, and the sectional areas of the inner cavities of the square cavity part, the transition cavity part and the circular cavity part are gradually reduced.
2. The 1536-well solid phase synthesis and extraction plate of claim 1, wherein the 1536 filter wells are arranged in a matrix of 48 equidistant filter wells in the transverse direction and 32 equidistant filter wells in the longitudinal direction.
3. The 1536-well solid phase synthesis and extraction plate of claim 1, wherein the cylinder assembly set comprises a first cylinder assembly chamber and a second cylinder assembly chamber, the first cylinder assembly chamber is disposed at the upper end of the second cylinder assembly chamber, and the cross-sectional diameter of the second cylinder assembly chamber is smaller than that of the first cylinder assembly chamber.
4. The 1536-well solid phase synthesis and extraction plate of claim 1, wherein the transition chamber portion has a tapered inner cross section along the direction from the square chamber portion to the circular chamber portion.
5. The 1536-well solid phase synthesis and extraction plate of claim 1, wherein the side length of the inner cavity of the cube assembly is not more than 3.7mm, and the height of the inner cavity is 0.1mm to 7 mm.
6. The 1536-well solid phase synthesis and extraction plate of claim 1, wherein the diameter of the inner cavity of the cylindrical assembly is not more than 3.5mm, and the height of the inner cavity is 0.1mm to 7 mm.
7. The 1536-well solid phase synthesis and extraction plate of claim 1, wherein the empty plate has a length ranging from 120mm to 520mm, a width ranging from 80mm to 360mm, and a height ranging from 10mm to 50 mm.
8. The 1536-well solid phase synthesis and extraction plate of claim 1, wherein the empty plate is provided with an identification block for indicating the placement direction of the empty plate.
9. The 1536-well solid phase synthesis and extraction plate of claim 1, wherein an X-axis code strip and a Y-axis code strip are respectively disposed on a surface of the hollow plate near the square inlet along X-axis and Y-axis directions.
10. The 1536-well solid phase synthesis and extraction plate of claim 1, wherein the outlet is in a bevel, or flat, or sharp configuration.
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