Kit for rapid extraction of plant seed DNA, and application thereof

A plant seed and kit technology, applied in the direction of DNA preparation, recombinant DNA technology, etc., can solve the problems of high impurities and low DNA yield, achieve high purity, improve yield and purity, and high yield

Active Publication Date: 2013-09-11
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of these extraction methods have the defects of low DNA yield and more impurities (containing more impurities such as proteins, lipids, polysaccharides, etc.), which need to be improved.

Method used

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  • Kit for rapid extraction of plant seed DNA, and application thereof
  • Kit for rapid extraction of plant seed DNA, and application thereof
  • Kit for rapid extraction of plant seed DNA, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0028] Experimental Example 1 pH value optimization experiment of column buffer

[0029] 1. Experimental materials

[0030] 1.1 Plant seeds: cotton seeds, rice seeds, corn seeds, soybean seeds.

[0031] 1.2 Extraction reagent: CTAB lysate: 1.4M NaCl, 2% CTAB, 0.1M Tris-HCl pH8.0, 0.02M EDTA;

[0032] Column-passing buffer: prepare 7 kinds of column-passing buffers, the components of which are the same: 5M guanidine hydrochloride, 0.05M Tris-HCl, 0.01M EDTA; the pH values ​​of the 7 column-passing buffers are 4.0 , 4.5, 5.0, 5.5, 6.0, 6.5, 7.0.

[0033] Rinsing solution 1: 3M NaAc pH5.2, absolute ethanol; wherein, the volume ratio of NaAc and absolute ethanol is 3:7;

[0034] Rinsing solution 2: 0.1M Tris-HCl pH7.0, absolute ethanol; wherein, the volume ratio of Tris-HCl and absolute ethanol is 3:7;

[0035] Elution buffer: TE buffer.

[0036] Silicon matrix membrane DNA adsorption column (purchased from Tomorrow Bio (Beijing) Technology Co., Ltd.);

[0037] 2. Experiment...

experiment example 2

[0070] The optimization experiment of guanidine hydrochloride concentration in the column buffer of experimental example 2

[0071] 1. Experimental materials

[0072] 1.1 Plant seeds: cotton seeds, rice seeds, corn seeds, soybean seeds.

[0073] 1.2 Extraction reagent:

[0074] CTAB lysate: 1.4M NaCl, 2% CTAB, 0.1M Tris-HCl pH8.0, 0.02M EDTA;

[0075] Column-passing buffer: 10 kinds of column-passing buffers were prepared, except for the concentration of guanidine hydrochloride, the other components of the 10 kinds of column-passing buffers were the same, namely: 0.05M Tris-HCl, 0.01M EDTA; The pH value is 5.5. The concentration of guanidine hydrochloride in the 10 column buffers is 2.5M, 3.0M, 3.5M, 4.0M, 4.5M, 5.0M, 5.5M, 6.0M, 6.5M, 7.0M;

[0076] Rinsing solution 1: 3M NaAc pH5.2, absolute ethanol; wherein, the volume ratio of NaAc and absolute ethanol is 3:7;

[0077] Rinsing solution 2: 0.1M Tris-HCl pH7.0, absolute ethanol; wherein, the volume ratio of Tris-HCl and ...

experiment example 3

[0113] Experimental example 3 The application experiment of kit of the present invention extracting plant seed DNA

[0114] 1. Experimental materials

[0115] 1.1 Plant seeds: cotton seeds, rice seeds, corn seeds, soybean seeds.

[0116] 1.2 Extraction reagent:

[0117]CTAB lysate: 1.4M NaCl, 2% CTAB, 0.1M Tris-HCl pH8.0, 0.02M EDTA;

[0118] Column passing buffer: 4.5M guanidine hydrochloride, 0.05M Tris-HCl, 0.01M EDTA; the pH of the column passing buffer is 5.5.

[0119] Rinsing solution 1: 3M NaAC pH5.2, absolute ethanol; wherein, the volume ratio of NaAC and absolute ethanol is 3:7;

[0120] Rinsing solution 2: 0.1M Tris-HCl pH7.0, absolute ethanol; wherein, the volume ratio of Tris-HCl and absolute ethanol is 3:7;

[0121] Elution buffer: TE buffer.

[0122] Silicon matrix membrane DNA adsorption column (Baiao Tomorrow (Beijing) Technology Co., Ltd.);

[0123] 2. Experimental method

[0124] (1) Weigh 100 mg each of corn, rice, and cotton seed powders, and 50 mg o...

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PUM

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Abstract

The invention discloses a kit for rapid extraction of plant seed DNA, and an application thereof. The compositions of the kit comprise: a CTAB lysate, a silicon-based plasmalemma DNA adsorption column, an RNA enzyme, a buffer solution for passing through a column, a rinsing solution 1, a rinsing solution 2 and an eluting buffer solution; the compositions of the buffer solution for passing through the column include: 2.5-7.0 M of guanidine hydrochloride, 0.05 M of Tris-HCl, and 0.01 M of EDTA; and the pH value is 4.0-7.0. The invention further discloses the application of the kit for the rapid extraction of the high quality plant seed DNA. The kit can effectively improve yield and purity of the extraction of the plant seed DNA, has the extracted DNA with good integrity, high purity and high yield; while decreasing DNA extraction cost, the kit effectively improves extraction quality and extraction efficiency of the plant seed genomic DNA.

Description

technical field [0001] The invention relates to a DNA extraction kit, in particular to a rapid plant seed DNA extraction kit and an application thereof, belonging to the field of plant seed DNA extraction. Background technique [0002] In molecular biology research, the extraction of high-quality DNA is the key to its downstream biological applications. With the development of molecular biology, DNA extraction methods of various plant materials have been established, including: CTAB method, SDS method, PVP method, urea method or high salt and low pH method, etc. The contents of proteins, polysaccharides, and phenolic substances in different plant materials vary greatly, and it is difficult to separate or purify high-quality DNA. In view of the differences in the cell walls and cell contents of different plant materials, the quality of DNA extraction can be guaranteed only by adopting an appropriate plant DNA extraction method. [0003] Plant seeds usually contain a large a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 金芜军张秀杰宛煜嵩贺辉群董美
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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