CN110819627A - Multi-column single tube consumable and use method thereof in nucleic acid extraction and purification by magnetic rod method - Google Patents
Multi-column single tube consumable and use method thereof in nucleic acid extraction and purification by magnetic rod method Download PDFInfo
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- CN110819627A CN110819627A CN201911205802.5A CN201911205802A CN110819627A CN 110819627 A CN110819627 A CN 110819627A CN 201911205802 A CN201911205802 A CN 201911205802A CN 110819627 A CN110819627 A CN 110819627A
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- 238000000034 method Methods 0.000 title claims abstract description 38
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 34
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 34
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 34
- 238000000605 extraction Methods 0.000 title claims abstract description 19
- 238000000746 purification Methods 0.000 title claims abstract description 15
- 239000011324 bead Substances 0.000 claims abstract description 57
- 238000005336 cracking Methods 0.000 claims abstract description 36
- 238000010828 elution Methods 0.000 claims abstract description 20
- 238000005406 washing Methods 0.000 claims abstract description 20
- 238000002156 mixing Methods 0.000 claims abstract description 12
- 238000000197 pyrolysis Methods 0.000 claims abstract description 8
- 239000000463 material Substances 0.000 claims abstract description 6
- 230000009089 cytolysis Effects 0.000 claims description 18
- 239000000243 solution Substances 0.000 claims description 14
- 238000013508 migration Methods 0.000 claims description 6
- 230000005012 migration Effects 0.000 claims description 6
- 239000012149 elution buffer Substances 0.000 claims description 4
- 239000011534 wash buffer Substances 0.000 claims description 4
- 230000001360 synchronised effect Effects 0.000 claims description 3
- 238000001179 sorption measurement Methods 0.000 abstract description 14
- 230000000694 effects Effects 0.000 abstract description 10
- 239000007788 liquid Substances 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
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- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
- C12N15/1013—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by using magnetic beads
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Abstract
The invention discloses a multi-column single-tube consumable and a use method thereof in nucleic acid extraction and purification by a magnetic rod method. The single tube consumptive material that current magnetic bead method nucleic acid extraction appearance used has restricted the further promotion of mixing and schizolysis effect. The multi-column single-tube consumable is provided with a cracking cavity and a plurality of functional cavities which are equally divided into two or three columns on the top surface, wherein each functional cavity comprises a washing cavity, an elution cavity and a magnetic bead placing cavity. According to the invention, as the space size of the pyrolysis cavity of the multi-column single-tube consumable is large, the high-frequency vibration of the sheath and the high-speed rotation of the operating platform can be combined to improve the uniform mixing and pyrolysis effects, and when the magnetic rod moves downwards to adsorb magnetic beads, the operating platform can rotate at a low speed, so that the magnetic rod is in contact with more space in the pyrolysis cavity, and the magnetic bead adsorption effect is improved; when the magnetic bar adsorbs the magnetic bead at the bottom of the cracking cavity, the operation platform rotates the cracking cavity to the coincidence of the diameter and the horizontal moving direction of the sheath, so that the motion track of the magnetic bar is longer, and the magnetic bead adsorption effect and the adsorption efficiency are improved.
Description
Technical Field
The invention belongs to the technical field of nucleic acid extraction and purification, and particularly relates to a multi-column single-tube consumable and a use method thereof in nucleic acid extraction and purification by a magnetic rod method.
Background
The existing single-tube consumables are all single-row consumables, and the size occupied along the consumable translation direction is too large, so that the size of a matched nucleic acid extractor along the consumable translation direction is also large; and the size of each washing cavity and elution cavity of the existing single-tube consumable along the translation direction of the vertical consumable cannot be designed to be very large, otherwise, the volume of each washing cavity and elution cavity is too large, and a large amount of washing buffer solution and elution buffer solution are needed to reach a certain height in the washing cavity and the elution cavity so as to meet the requirements during washing and elution. In addition, the length dimension of single-row single-tube consumables is great, and the space that takes up length direction is great for accomodating range upon range of, causes inconvenience for accomodating.
The magnetic bead method nucleic acid extractor is a common nucleic acid extractor, and the magnetic bead method nucleic acid extractor adopts a magnetic rod method for nucleic acid extraction most commonly. The magnetic rod method is to realize the separation of nucleic acid by fixing liquid and transferring magnetic beads, separate the magnetic beads from waste liquid by the adsorption of the magnetic rods on the magnetic beads, and put the separated magnetic beads into the liquid in the next step to realize the extraction of nucleic acid, which usually comprises the processes of cracking, adsorption, washing and elution. However, when cracking, because the single-tube consumables adopted by the existing magnetic bead method nucleic acid extraction instrument are all single-row, the size perpendicular to the translation direction of the consumables is limited, if the single-tube consumables are subjected to horizontal auxiliary vibration to realize better uniform mixing and cracking of the reagent, the horizontal auxiliary vibration track is limited, and the uniform mixing and cracking effects are to be improved.
In addition, adopt single-column single tube consumptive material, when the bar magnet moved down in the schizolysis chamber and carried out the magnetic bead and adsorb, because the space size of single tube consumptive material is limited, the bar magnet only is fit for the motion in a direction of vertical direction, even also carry out the translation in the horizontal direction, the displacement of translation is also very little, and is visible, and the magnetic bead adsorption effect is relatively poor.
Disclosure of Invention
The invention aims to provide a multi-column single-tube consumable and a use method thereof in nucleic acid extraction and purification by a magnetic rod method, aiming at the defects of the prior art.
According to the multi-column single-tube consumable, the top surface is provided with the cracking cavity and the plurality of functional cavities, the functional cavities are divided into two or three columns which are arranged side by side, and each functional cavity comprises the washing cavity, the elution cavity and the magnetic bead placing cavity.
Preferably, the lysis chamber is cylindrical or rectangular.
Preferably, the washing cavity, the elution cavity or the magnetic bead placing cavity has a cuboid upper half section and a cylindrical lower half section, or is integrally a cylindrical cavity.
The invention relates to a use method of multi-column single-tube consumables in nucleic acid extraction and purification by a magnetic rod method, which comprises the following steps:
uniformly mixing: sucking the magnetic beads in the magnetic bead placement cavity and injecting the magnetic beads into the cracking cavity; the sheath is sleeved into the cracking cavities of the multiple rows of single-tube consumables, and the sheath starts to vibrate; meanwhile, the operating platform rotates according to the set plane motion track, so that the multiple rows of single-tube consumables are driven to rotate.
Magnetic attraction: after the sample to be detected, the lysis solution and the magnetic beads in the lysis cavity are mixed uniformly, stopping the operation platform; the sheath is moved out of the cracking cavities of the multiple rows of single-tube consumables, and the magnetic rod is inserted into the sheath; then, the operation platform rotates at a lower rotating speed than that during mixing, the sheath and the magnetic rod are synchronously sleeved in the cracking cavities of the multiple rows of single-tube consumables, and magnetic beads with nucleic acid adsorbed in the cracking cavities are adsorbed layer by layer; after sheath and bar magnet reached the bottom position in schizolysis chamber, operation platform rotated the horizontal migration direction coincidence of its diameter and sheath with the schizolysis chamber, then sheath and bar magnet synchronous horizontal migration adsorb the magnetic bead that the bottom of schizolysis chamber had the nucleic acid.
Preferably, the planar motion track is circular or elliptical.
Preferably, the distance between the central point of the lysis cavity of the multi-column single-tube consumable and the central point of the plane motion trajectory of the lysis cavity is equal to or greater than 1/2 of the central distance between two adjacent functional cavities.
Preferably, prior to blending: n multi-column single tube consumable sets are arranged on the operation platform at intervals, each multi-column single tube consumable set comprises m multi-column single tube consumables, n is more than or equal to 3 and less than or equal to 6, and m is more than or equal to 3 and less than or equal to 6; the multiple rows of single tube consumables are fixed with the operating platform; injecting a sample to be detected and lysis solution into the lysis cavity of the multi-column single-tube consumable, and injecting washing buffer solution, elution buffer solution and magnetic beads into the washing cavity, the elution cavity and the magnetic bead placing cavity respectively.
Preferably, the multi-column single tube consumables and the operation platform are fixed in a manner that four positioning holes of the multi-column single tube consumables or two positioning holes at an oblique angle position are positioned through a cylinder fixed on the operation platform, and the multi-column single tube consumables are fastened on the operation platform through a buckle.
The invention has the beneficial effects that:
1. the invention designs a multi-column single-tube consumable, which can reduce the size of a matched nucleic acid extractor along the translation direction of the consumable. The advantages of the magnetic rod method nucleic acid extraction and purification by using the multi-column single-tube consumable material of the invention are as follows: because the pyrolysis cavity space size of multiseriate single tube consumptive material is great, when the sheath was in the pyrolysis cavity high-frequency oscillation from top to bottom, operation platform can be according to the high-speed rotary motion of the planar motion orbit that sets for, and mixing and pyrolysis effect can be improved greatly in two motion combinations. In addition, when the magnetic rod and the sheath move downwards together in the cracking cavity for magnetic bead adsorption, the operating platform can rotate at a low speed, so that the magnetic rod is in contact with more spatial positions in the cracking cavity when adsorbing magnetic beads with nucleic acid in the cracking cavity layer by layer, and the magnetic bead adsorption effect and the adsorption efficiency are greatly improved; when the magnetic bar and the sheath reach the bottom of the cracking cavity together for magnetic bead adsorption, the operation platform enables the cracking cavity to rotate to the position where the diameter of the cracking cavity coincides with the horizontal moving direction of the sheath, so that the motion track of the magnetic bar and the sheath is long, and the magnetic bead adsorption effect and the adsorption efficiency are improved. Therefore, the magnetic rod method nucleic acid extraction and purification can be carried out by using the multi-column single-tube consumable, and the magnetic bead adsorption effect and the magnetic bead adsorption efficiency can be improved from multiple aspects.
2. The multi-row single tube consumable is moderate in length-width ratio, and the space occupied by the storage and stacking in the length direction and the width direction is balanced, so that the storage is facilitated.
Drawings
FIG. 1 is a perspective view of the structure of the present invention;
fig. 2 is a top view of the structure of the present invention.
Detailed Description
The invention is further explained with reference to the drawings and the embodiments.
As shown in FIG. 1, the present embodiment is a dual-row single-tube consumable, which has a top surface provided with a lysis chamber 1 and six functional chambers, wherein the six functional chambers are equally divided into two rows arranged side by side, and the six functional chambers are respectively four washing chambers, an elution chamber and a magnetic bead placement chamber; as a preferred embodiment, the lysis chamber is numbered 1; the two function cavities closest to the cracking cavity are numbered 2 and 5, and the function cavity with the serial number 5 and the function cavity with the serial number 2 are arranged along the outer edge of the cracking cavity in a counterclockwise way; the same row of function cavities with serial number 2 and the serial number of the adjacent function cavity is marked as 3, and the serial number of the remaining function cavity in the same row with the function cavity with serial number 3 is marked as 4; the same row of function cavities with serial number 5 and the serial number of the adjacent function cavity is marked as 6, and the serial number of the remaining function cavity in the same row with the function cavity with serial number 6 is marked as 7; the functional cavity with the serial number of 4 is used as a magnetic bead placing cavity, the functional cavity with the serial number of 7 is used as an elution cavity, the rest four functional cavities are used as washing cavities, and of course, the functional cavities with other serial numbers can be used as an elution cavity and a magnetic bead placing cavity; as a preferred embodiment, the lysis chamber may have a cylindrical or rectangular parallelepiped shape, and the washing chamber, the elution chamber, or the magnetic bead placement chamber may have a rectangular parallelepiped shape in the upper half and a cylindrical shape in the lower half, or may have a cylindrical chamber as a whole.
The use method of the double-row single tube consumable in nucleic acid extraction and purification by a magnetic rod method comprises the following specific steps:
n double-row single-tube consumable sets are arranged on the operating platform at intervals, each double-row single-tube consumable set comprises m double-row single-tube consumables, n is more than or equal to 3 and less than or equal to 6, and m is more than or equal to 3 and less than or equal to 6; four positioning holes of the double-row single-tube consumable or two positioning holes at the oblique angle position are positioned through a cylinder fixed on the operation platform, and the double-row single-tube consumable is fastened on the operation platform through a buckle; injecting a sample to be detected and lysis solution into a lysis cavity of the double-row single-tube consumable, and respectively injecting washing buffer solution, elution buffer solution and magnetic beads into a washing cavity, an elution cavity and a magnetic bead placing cavity; then, sucking the magnetic beads in the magnetic bead placing cavity and injecting the magnetic beads into the cracking cavity; the sheath is sleeved in a cracking cavity of the double-row single-tube consumable, and the sheath starts to vibrate; meanwhile, the operation platform is designed to be circular or elliptical according to a set plane motion track (in the embodiment, the plane motion track of the operation platform is specifically designed to be circular or elliptical, and a motor is used for driving an eccentric wheel to deflectThe eccentric wheel drives the operating platform to move synchronously) to rotate, thereby driving the double-row single-tube consumables to rotate; wherein, the distance L between the center point of the cracking cavity of the double-row single-tube consumable and the center point of the plane motion track of the cracking cavity1Equal to or more than the center distance L between two adjacent functional cavities 21/2, as shown in fig. 2. After the sample to be detected, the lysis solution and the magnetic beads in the lysis cavity are mixed uniformly, stopping the operation platform; the sheath is moved out of a cracking cavity of the double-row single-tube consumable, and the magnetic rod is inserted into the sheath; then, the operation platform rotates at a lower rotating speed than that during mixing, the sheath and the magnetic rod are synchronously sleeved in a cracking cavity of the double-row single-tube consumable, and magnetic beads with nucleic acid adsorbed in the cracking cavity are adsorbed layer by layer; after sheath and bar magnet reached the bottom position in schizolysis chamber, operation platform rotated the horizontal migration direction coincidence of its diameter and sheath with the schizolysis chamber, then sheath and bar magnet synchronous horizontal migration adsorb the magnetic bead that the bottom of schizolysis chamber had the nucleic acid. After the magnetic beads at the bottom of the cracking cavity are adsorbed, the sheath and the magnetic rod synchronously move out of the cracking cavity of the double-row single-tube consumable and are synchronously sleeved in the washing cavity of the double-row single-tube consumable to wash nucleic acid, and after the four washing cavities are washed, the sheath and the magnetic rod are synchronously sleeved in the elution cavity of the double-row single-tube consumable to elute the nucleic acid.
Claims (8)
1. Multiseriate single tube consumptive material has seted up the schizolysis chamber at the top surface, its characterized in that: still seted up a plurality of function chambeies at the top surface, a plurality of function chambeies are equallyd divide for two or three rows that set up side by side, and the function chamber contains washing chamber, elution chamber and magnetic bead and places the chamber.
2. The multi-column single tube consumable of claim 1, wherein: the cracking cavity is cylindrical or cuboid.
3. The multi-column single tube consumable of claim 1, wherein: the upper half section of the washing cavity, the elution cavity or the magnetic bead placing cavity is cuboid, the lower half section of the washing cavity, the elution cavity or the magnetic bead placing cavity is cylindrical, or the whole washing cavity, the elution cavity or the magnetic bead placing cavity is a cylindrical cavity.
4. The use method of the multi-column single-tube consumable in nucleic acid extraction and purification by a magnetic rod method according to any one of claims 1 to 3, wherein the method comprises the following steps: the method comprises the following specific steps:
uniformly mixing: sucking the magnetic beads in the magnetic bead placement cavity and injecting the magnetic beads into the cracking cavity; the sheath is sleeved into the cracking cavities of the multiple rows of single-tube consumables, and the sheath starts to vibrate; meanwhile, the operating platform rotates according to a set plane motion track, so that a plurality of rows of single-tube consumables are driven to rotate;
magnetic attraction: after the sample to be detected, the lysis solution and the magnetic beads in the lysis cavity are mixed uniformly, stopping the operation platform; the sheath is moved out of the cracking cavities of the multiple rows of single-tube consumables, and the magnetic rod is inserted into the sheath; then, the operation platform rotates at a lower rotating speed than that during mixing, the sheath and the magnetic rod are synchronously sleeved in the cracking cavities of the multiple rows of single-tube consumables, and magnetic beads with nucleic acid adsorbed in the cracking cavities are adsorbed layer by layer; after sheath and bar magnet reached the bottom position in schizolysis chamber, operation platform rotated the horizontal migration direction coincidence of its diameter and sheath with the schizolysis chamber, then sheath and bar magnet synchronous horizontal migration adsorb the magnetic bead that the bottom of schizolysis chamber had the nucleic acid.
5. The use method of the multi-column single-tube consumable in nucleic acid extraction and purification by the magnetic rod method according to claim 4, wherein the method comprises the following steps: the plane motion trail is circular or elliptical.
6. The use method of the multi-column single-tube consumable in nucleic acid extraction and purification by the magnetic rod method according to claim 4, wherein the method comprises the following steps: the distance between the central point of the pyrolysis cavity of the multi-column single-tube consumable and the central point of the plane motion track of the pyrolysis cavity is equal to or more than 1/2 of the central distance between two adjacent functional cavities.
7. The use method of the multi-column single-tube consumable in nucleic acid extraction and purification by the magnetic rod method according to claim 4, wherein the method comprises the following steps: before uniformly mixing: n multi-column single tube consumable sets are arranged on the operation platform at intervals, each multi-column single tube consumable set comprises m multi-column single tube consumables, n is more than or equal to 3 and less than or equal to 6, and m is more than or equal to 3 and less than or equal to 6; the multiple rows of single tube consumables are fixed with the operating platform; injecting a sample to be detected and lysis solution into the lysis cavity of the multi-column single-tube consumable, and injecting washing buffer solution, elution buffer solution and magnetic beads into the washing cavity, the elution cavity and the magnetic bead placing cavity respectively.
8. The use method of the multi-column single-tube consumable in nucleic acid extraction and purification by the magnetic rod method according to claim 7, characterized in that: the multi-column single tube consumable is fixed on the operating platform in a mode that four positioning holes of the multi-column single tube consumable or two positioning holes at oblique angle positions are positioned through a cylinder fixed on the operating platform, and the multi-column single tube consumable is fastened on the operating platform through a buckle.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201911205802.5A CN110819627A (en) | 2019-11-29 | 2019-11-29 | Multi-column single tube consumable and use method thereof in nucleic acid extraction and purification by magnetic rod method |
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| CN201911205802.5A CN110819627A (en) | 2019-11-29 | 2019-11-29 | Multi-column single tube consumable and use method thereof in nucleic acid extraction and purification by magnetic rod method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111334417A (en) * | 2020-04-14 | 2020-06-26 | 无锡科智达科技有限公司 | Pre-packaging magnetic bead method nucleic acid extraction device |
| CN117757786A (en) * | 2024-02-22 | 2024-03-26 | 山东艾克韦生物技术有限公司 | nucleic acid purification auxiliary device and nucleic acid purification experiment method |
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2019
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