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
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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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