Low-concentration PCR product concentrating method used for direct sequencing
The invention relates to a low-concentration PCR product concentrated perborate and 2ul nucleic acid settling agent (artificially synthesized DNA sequence), and mixing evenly; adding a 2-2 method. The 2-2 method comprises the specific steps of adding 3M sodium acetate with 1/10 volume and pH of 5.2,...
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| creator | QING ZHIRONG TIAN QING |
| description | The invention relates to a low-concentration PCR product concentrated perborate and 2ul nucleic acid settling agent (artificially synthesized DNA sequence), and mixing evenly; adding a 2-2 method. The 2-2 method comprises the specific steps of adding 3M sodium acetate with 1/10 volume and pH of 5.2, absolute ethyl alcohol with 0.5 volume and 300 ul 40% PEG8000 into a PCR product to be purified, placing the solution at -20 DEG C for 10-30 min; conducting centrifugation at 4 DEG C and 12000 rpm for 10-20 min, and abandoning the supernatant; adding 1 ml 70% ethyl alcohol to be subjected to precooling at 4 DEG C, reversing evenly, conducting centrifugation at 4 DEG C and 12000 rpm for 2-10 min, and abandoning the supernatant; placing a centrifuge tube with a cover opened at room temperature, and making the remaining liquid fully volatilize; adding water or a TE solution to dissolve precipitated DNA. By the adoption of the method, the recycled product can be directly used for sequencing, the success rate of the lo |
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The 2-2 method comprises the specific steps of adding 3M sodium acetate with 1/10 volume and pH of 5.2, absolute ethyl alcohol with 0.5 volume and 300 ul 40% PEG8000 into a PCR product to be purified, placing the solution at -20 DEG C for 10-30 min; conducting centrifugation at 4 DEG C and 12000 rpm for 10-20 min, and abandoning the supernatant; adding 1 ml 70% ethyl alcohol to be subjected to precooling at 4 DEG C, reversing evenly, conducting centrifugation at 4 DEG C and 12000 rpm for 2-10 min, and abandoning the supernatant; placing a centrifuge tube with a cover opened at room temperature, and making the remaining liquid fully volatilize; adding water or a TE solution to dissolve precipitated DNA. 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| subjects | BEER BIOCHEMISTRY CHEMISTRY COMPOSITIONS OR TEST PAPERS THEREFOR COMPOSITIONS THEREOF CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES CULTURE MEDIA ENZYMOLOGY MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS METALLURGY MICROBIOLOGY MICROORGANISMS OR ENZYMES MUTATION OR GENETIC ENGINEERING PROCESSES OF PREPARING SUCH COMPOSITIONS PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS SPIRITS VINEGAR WINE |
| title | Low-concentration PCR product concentrating method used for direct sequencing |
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