Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for reclaiming DNA from agarose gel

An agarose gel and electrophoresis technology, applied in the field of DNA recovery and purification, can solve the problems of easy cross-contamination of multiple samples recovered by cutting gel, and achieve the effects of eliminating gel cutting steps, overcoming cumbersome operations, and easy operation.

Inactive Publication Date: 2008-10-01
GUIYANG MEDICAL UNIVERSITY
View PDF1 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to overcome the existing method, the cumbersome operation of gel cutting recovery and the shortcoming of easy cross-contamination when operating multiple samples, and to provide a method for reclaiming linear or circular DNA from agarose gel after electrophoresis, This method can recover and purify the target DNA fragment from the agarose gel within 15 minutes, and the purity is very high, which can meet the requirements of subsequent molecular cloning, restriction analysis, labeling and PCR.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] DNA was recovered from the agarose gel after electrophoresis as follows:

[0019] 1) Cut the hard water-absorbing sponge that has been sterilized in advance into 3mm×3mm×10mm, and the lower end is a wedge-shaped strip.

[0020] 2) After electrophoresis, align the wedge-shaped hard water-absorbing sponge strip with the target DNA band and insert it into the gel under the irradiation of ultraviolet light. After it is absorbed for 20 seconds, it can be seen that the fluorescent DNA band is fully absorbed. Breathe it in, then pull it out.

[0021] 3) Put it into a centrifuge tube with a funnel and centrifuge at 15000rpm for 3 minutes to centrifuge the DNA solution from it.

[0022] 4) Place the DNA recovery column in a centrifuge tube, transfer the DNA solution onto the DNA recovery column, place at room temperature for 2 minutes, centrifuge at 12,000 rpm for 1 minute, and discard the waste liquid at the bottom of the tube.

[0023] 5) Add 100 μl of DNA washing solution t...

Embodiment 2

[0027] Recover plasmid DNA (closed circular DNA) from the agarose gel after electrophoresis according to the following procedure:

[0028] 1) Cut the hard water-absorbing sponge that has been sterilized in advance into 3mm×3mm×10mm, and the lower end is a wedge-shaped strip.

[0029] 2) After the electrophoresis is completed, align the wedge-shaped hard water-absorbing sponge strip with the target plasmid DNA band under the irradiation of ultraviolet light and insert it into the gel. After it is absorbed for 20 seconds, the fluorescent DNA band can be seen to be absorbed. Inhale it fully, then pull it out.

[0030] 3) Put it into a centrifuge tube with a funnel and centrifuge at 15,000 rpm for 5 minutes to centrifuge the plasmid DNA solution therefrom. It can be used directly for subsequent experiments, or stored at -20°C for future use.

[0031] In this example, the DNA sample recovered from the agarose gel within 6 minutes, A 260 / A 280 It is 1.45, which can meet the sub...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a method for recycling DNA from agarose gel. The method for recycling DNA samples from agarose gel after electrophoresis comprises the following steps in sequence: first, stripped absorbent material of wedge type is prepared, then inserted into gel, sucks out the DNA samples, and then the method is finished through centrifugal collection, DNA absorption, DNA washing, DNA elution, etc; the invention overcomes the defects existing in the prior art of complicated operation of cutting gel for recycling and easy cross contamination when operating a plurality of samples, provides a method for recycling liner or annular DNA from agarose gel after electrophoresis, can recycle DNA segments for purification from agarose gel within 15min, has high pureness and can meet the subsequent operation of molecular cloning, enzyme analysis, marking, PCR, etc.; the method adopts easy equipment, has convenient operation, high recycling rate and low cost and can put reagent in kits.

Description

technical field [0001] The patent of the present invention relates to a method for recovering and purifying DNA, in particular to a method for recovering and purifying DNA from agarose gel after electrophoresis. Background technique [0002] In the field of molecular biology technology, many experiments need to separate DNA fragments by agarose electrophoresis, and then recover the required DNA bands for subsequent experiments, such as DNA fragment cloning, enzyme digestion analysis, labeling, polymerase chain reaction ( polymerase chain reaction, abbreviated as PCR) and in vitro transcription. The recovery of agarose gel DNA has always been the main content of molecular biology experiments. Since agarose gel is solid, the DNA bands contained in it are not easy to be released, so its recovery has always required multi-step operations. [0003] There are many Chinese invention patent applications related to the DNA field, the application number is 200610037351.5, and the inv...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07H21/04C07H1/06
Inventor 黄海冯发莉刘杨黄健
Owner GUIYANG MEDICAL UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com