Marker for auxiliary detection of wheat high SNP (Single Nucleotide Polymorphism) and application thereof
An auxiliary detection and spikelet number technology, which is applied in the field of wheat genetics and breeding, can solve the problems of poor stability, heavy workload, and low efficiency, and achieve the effects of stable labeling, easy operation, and mature and stable technology
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[0027] The present invention will be described in detail below in combination with specific embodiments.
[0028] 1. Using CTAB method to extract wheat genomic DNA, extraction steps:
[0029] (1) Take 100 mg of fresh young leaves, grind them into fine powder with liquid nitrogen, add 2×CTAB extract (2% CTAB; 1.4M NaCl, 0.1MTris-HCl, PH8.0, 0.1M EDTA) preheated to 65°C ,PH8.0) 1ml, mix well;
[0030] (2) 65 ℃ water bath for 30-45min, during which shake gently to mix. After cooling to room temperature, add an equal volume of chloroform:isoamyl alcohol (24:1), mix gently until the supernatant becomes milky, and centrifuge at 4000rpm for 10min;
[0031] (3) Take the supernatant, add an equal volume of isopropanol, and place in an ice bath to precipitate DNA;
[0032] (4) Centrifuge at 4000rpm for 10min, remove the supernatant, wash twice with 70% ethanol and once with absolute ethanol. Air-dry the DNA and dissolve it in an appropriate amount of 1×TE solution with pH 8.0. Add R...
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