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Vibration eluting method for improving recovery efficiency of DNA recycling reagent kit

A technology of recovery efficiency and elution method, which is applied in the field of DNA recovery kits, can solve the problems of affecting the progress of the experiment, wasting time and reagents, and light DGGE bands, etc., and achieve the effect of increasing the concentration of DNA recovery, improving the effect and working efficiency

Inactive Publication Date: 2011-04-20
XIAMEN UNIV +1
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Problems solved by technology

[0009] Such recovery results often have a low DNA concentration, and some samples often fail to meet the requirements of subsequent experiments. For example, for DGGE experiments or sequencing, PCR and gel recovery are required, which wastes reagents and time, and affects the progress of the experiment.
Some samples that can barely be used for follow-up experiments, due to the low concentration of DNA, also affect the experimental results, such as DGGE bands are light and fuzzy, some information cannot be expressed, and DGGE experiments need to be repeated, which is a waste of time and reagents

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  • Vibration eluting method for improving recovery efficiency of DNA recycling reagent kit
  • Vibration eluting method for improving recovery efficiency of DNA recycling reagent kit

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[0025] The present invention improves step 5) in the existing recovery method (see the experimental steps of the kit instruction manual): only add 1 eluent to the column, and the amount of eluent added is according to the one-time addition amount of the recovery kit According to the general recovery kit, the addition amount is 25-50 μl, preferably 30 μl, let it stand for 0-5 minutes, preferably after 1 minute, insert the centrifuge tube with the column into the hole of the shaker, and use 500 ~1000rpm / min, preferably 800rpm / min, according to the test results, such as figure 1 As shown, the shaking time is preferably 0.5-5 minutes, preferably 2 minutes, and the recovery concentration of DNA is the largest, which is about 20% higher than that without shaking.

[0026] The result after improvement was compared with the result before improvement, that is, the result of only adding 30 μl of eluent once and shaking, compared with the method of adding 25 μl in two times according to ...

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Abstract

An oscillation eluting method used for improving the recovery efficiency of a DNA extraction kit relates to a DNA extraction kit, in particular to an oscillation method added in the operation steps of the DNA extraction kit, so as to improve the recovery efficiency of DNA and be used for PCR product recycling of DNA segments. The invention provides an oscillation eluting method which has higher recovery efficiency, can meet the subsequent experimental requirements and improve the recovery efficiency of a DNA extraction kit. The invention has the technical proposal that on the basis of the existing recovery method of the DNA extraction kit, the step 5 is improved, eluent is only needed to be added in a pillar for once, and the quantity of the eluent is counted according to the adding quantity of the extraction kit for once; after the eluent stands still, a centrifugal tube provided with the pillar is inserted into a hole of an oscillator and is vibrated at the speed of 500-1000rpm / min,and the DNA is recovered. The recycling concentration of the vibrated eluent reaches the maximum value which is increased by about 20% compared with the concentration of the eluent which is not vibrated.

Description

technical field [0001] The invention relates to a DNA recovery kit, in particular to a method for adding a shock to the operation steps of the DNA recovery kit to improve the recovery efficiency of DNA and to recover PCR products of DNA fragments. Background technique [0002] DNA recovery kits produced by foreign companies (such as Takara) generally have high recovery efficiency, but they are expensive and the experimental steps are cumbersome. The cost of reagents for recovering each sample is about 5 times that of domestic kits (such as Biotech). Domestic recovery kits are cheap and easy to operate, but the recovery efficiency is low, which affects subsequent experiments, such as denaturing gradient gel electrophoresis (DGGE) experiments, DNA fragment sequencing, etc. Many domestic scholars have discussed methods to improve the recovery efficiency of DNA, mainly electrophoresis ([1] Jiang An, Liang Hui, Chen Xu, etc. A simple method for recovering DNA in agarose gel. Biot...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
Inventor 汤坤贤焦念志
Owner XIAMEN UNIV
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