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Sol buffer for gel DNA recovery, kit for gel DNA recovery and application method of kit

A kit and gel technology, applied in the field of DNA recovery, can solve the problems of unstable supporting sol buffer, long DNA recovery time of gel, and limited DNA recovery capacity, etc., and achieve the effects of low cost, efficient recovery, and simple preparation method

Inactive Publication Date: 2018-08-28
爱默生物科技(厦门)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The invention mainly solves the problems of long gel DNA recovery time, high economic cost, limited DNA recovery capacity, unstable sol buffer matched with Glass milk, etc.

Method used

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  • Sol buffer for gel DNA recovery, kit for gel DNA recovery and application method of kit
  • Sol buffer for gel DNA recovery, kit for gel DNA recovery and application method of kit
  • Sol buffer for gel DNA recovery, kit for gel DNA recovery and application method of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Example 1: Determination of the composition of the supporting sol buffer for Glass milk

[0077] Select an equal amount of the same DNA sample (PBOBI plasmid DNA, the size is about 9000bp, can be purchased from Protin Biotechnology (Beijing) Co., Ltd.), after 1% agarose gel electrophoresis, cut the DNA band, the size of the cut strip is basically In the same way, sol buffers with different components were added (Buffer A: 3M GuSCN+3M NaCl; Buffer B: 3M GuSCN+3M NaAC; Buffer C: 3M GuSCN+3M GuHCl; Buffer D: 6M GuSCN). Gel DNA recovery step for DNA recovery, and finally use an equal volume of sterile water for elution, take an equal volume of DNA solution for 1% agarose electrophoresis detection, the results are shown in figure 1 . Among them, the 3M GuSCN+3M NaCl lane indicates that the sol buffer uses 3M GuSCN+3MNaCl; the lane 3M GuSCN+3M NaAC indicates that the sol buffer uses 3M GuSCN+3M NaAC; buffer C: the lane 3M GuSCN+3M GuHCl indicates that the sol The buffer use...

Embodiment 2

[0078] Example 2: Optimization of the concentration of each component of the sol buffer

[0079] Select the same amount of the same DNA sample, cut the DNA band after 1% agarose gel electrophoresis, the size of the cut gel strip is basically the same, use the mixed solution of different concentrations of GuSCN and NaCl (Buffer A: 3M GuSCN+3M NaCl; Buffer B : 3.2M GuSCN+3M NaCl; Buffer C: 3M GuSCN+2M NaCl; Buffer D: 3.2M GuSCN+2M NaCl; Buffer E: 3M GuSCN+1.5M NaCl; Buffer F: 3.2M GuSCN+1.5M NaCl) as a sol buffer, use the gel DNA recovery step in this study to recover DNA, and finally use an equal volume of sterile water for elution, then take an equal volume of DNA solution for 1% agarose electrophoresis detection, the results are shown in figure 2 . The lane 3M GuSCN+3M NaCl means that the sol buffer is 3M GuSCN+3M NaCl; the lane 3.2M GuSCN+3M NaCl means that the sol buffer is 3.2M GuSCN+3M NaCl; the lane 3M GuSCN+2M NaCl means the sol The buffer uses 3M GuSCN+2MNaCl; the 3...

Embodiment 3

[0081] Example 3: Comparison of the ability of commonly used sol buffers to cooperate with Glass milk (that is, DNA-specific adsorption materials) to recover DNA

[0082] The Glass milk prepared by this scheme is combined with AG buffer, sol buffer—QG buffer in the Qiagen kit, and 6M NaI for gel DNA recovery. Finally, an equal volume of sterile water is used for elution, and an equal amount of DNA solution is used for elution. After 1% agarose electrophoresis detection, the results are shown in image 3 and 4 . Among them, the label AG buffer indicates that the sol buffer (3.2M guanidine isothiocyanate and 3M NaCl) of the present invention is used, the label QG buffer indicates that the sol buffer in the Qiagen kit is used, and the label 6M NaI indicates that the 6M NaI is used. NaI is used as sol buffer. Traditionally, the sol buffer used with Glass milk is 6M NaI. Each buffer was repeated three times independently.

[0083] Depend on image 3 The three lanes correspond...

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Abstract

The invention discloses a sol buffer for gel DNA recovery, a kit for gel DNA recovery and an application method of the kit. The sol buffer is prepared from guanidinium isothiocyanate and NaCl, and preferably from 3-3.2M guanidinium isothiocyanate and 1.5-3M NaCl. The invention also discloses a preparation method of a DNA specific adsorption material matching the sol buffer, and a kit for gel DNA recovery and an application method of the kit. The sol buffer for gel DNA recovery and the matching DNA specific adsorption material are of low cost, the preparation is simple and quick (about 2 hours), the application is easy, the effect is good, and the recovery rate is as high as 50% and above; simply by adopting conventional instrument and reagents of a laboratory, the initial amount of singleDNA recovery can reach 200mu g and above, and the recovered DNA can be applied to follow-up PCR, sequencing, etc.

Description

technical field [0001] The invention relates to the field of DNA recovery, in particular to a sol buffer for recovering gel DNA, a kit for recovering gel DNA and a use method thereof. Background technique [0002] In the process of gene cloning experiments, purification and recovery of DNA after agarose electrophoresis is the most common experimental link. At present, common methods for DNA purification and recovery after agarose electrophoresis include: low melting point agarose gel recovery, kit column recovery and glass milk method. However, these methods are relatively cumbersome, time-consuming and costly per experiment. In addition, for the glass milk DNA recovery method, the main component of the sol buffer used is NaI, which is easily oxidized by oxygen in the air and precipitates and changes color, which affects DNA recovery. [0003] At present, the most commonly used gel DNA recovery method in the laboratory is the kit column recovery method. Such as Qiaquick G...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/10
Inventor 梁耀极刘天星陈莹庄明火徐国标赵燕燕潘旭鲁靖
Owner 爱默生物科技(厦门)有限公司
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