Molecule mark identification method for plum pollen autocopulation non-compatible genotype
A technology of molecular markers and identification methods, applied in biochemical equipment and methods, measurement/inspection of microorganisms, DNA/RNA fragments, etc., can solve the problems that pollen S genotype identification cannot be carried out, and achieve simple identification methods and easy operation methods Simple steps and the effect of saving production costs
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[0024] 1. Refer to the method of Doyle and Doyle (Doyle JI, DoyleJL. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull, 1987, 19: 11-15) to extract the genomic DNA of 11 plum varieties: banana plum, royal Diamond, 82-1-109, Santa rosa ShSk, Younai, Wixon, Rose Plum, Nvgelei, Huangpi Plum, Black Jewel, Gaixian Dali, all varieties come from the National Germplasm Bank of Xingli (Liaoning Xiongyue)".
[0025] 2. By comparing the conserved regions of cherry, plum, almond and apricot pollen SFB genes, design general primers for plum pollen SFB gene according to the conserved sequence:
[0026] Forward primer 5′-TTKSCHATTRYCAACCKCAAAAG-3′
[0027] Reverse primer 5′-CAMRAATTCGATTTCGYCATATTTC-3′
[0028] The plum genomic DNA was amplified with the designed universal primers.
[0029] Reaction system: 25μL total reaction volume, 10×PCR Buffer 2.5μL 3.0mM MgCl 2 , 0.2mM dNTP, each primer 0.1μM, 20-50ng template, Taq enzyme 1.0U.
[0030] The ...
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