Molecular marker primer composition for identifying freestone trait of prunus persica fruits and application of molecular marker primer composition
A primer combination and molecular labeling technology, applied in the field of molecular biology, can solve the problems that the last single base cannot achieve the resolution effect and the primer cannot be effectively extended, and achieve the effect of reducing the test cost, reliable detection effect and cost reduction.
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Embodiment 1
[0047] A set of application methods for identifying the molecular marker primer combination of peach sticky core traits, the detailed steps are:
[0048] (1) Primer synthesis: According to the sequence information in Table 1, Sangon Bioengineering (Shanghai) Co., Ltd. was entrusted to synthesize primers JYSNP-1, JYSNP-2, JYSNP-3 and tail primers respectively; wherein, the 5' end of the tail primers Add three different fluorophores, namely FAM, HEX and NED;
[0049] Table 1: Related Primer Sequences
[0050]
[0051] Note: * indicates thio modification.
[0052] (2) Extraction of peach leaf DNA of the population sample to be tested:
[0053] A. Grind about 0.2g of young leaves with liquid nitrogen into powder and place them in a 2mL centrifuge tube;
[0054] B. Add 0.8mL CTAB lysate preheated at 65°C and mix well. The formula of the lysate is: 100mmol / L Tris Cl (pH 8.0), 20mmol / L EDTA (pH 8.0), 1.4mmol / L NaCl, 2.5% CTAB (M / V), 3% PVP (M / V), 2% β-mercapto...
Embodiment 2
[0067] A set of application methods for identifying the molecular marker primer combination of peach sticky core traits, the detailed steps are:
[0068] (1) Primer synthesis: According to the sequence information in Table 1, entrust a relevant biotechnology company to synthesize primers JYSNP-1, JYSNP-2, and JYSNP-3 and choose one of them (only primer JYSNP-2 was synthesized in this example);
[0069] Table 1: Related Primer Sequences
[0070]
[0071] Note: * indicates thio modification.
[0072] (2) Extraction of peach leaf DNA of the population sample to be tested:
[0073] A. Grind about 0.2g of young leaves with liquid nitrogen into powder and place them in a 2mL centrifuge tube;
[0074] B. Add 0.8mL CTAB lysate preheated at 65°C and mix well. The formula of the lysate is: 100mmol / L Tris Cl (pH 8.0), 20mmol / L EDTA (pH 8.0), 1.4mmol / L NaCl, 2.5% CTAB (M / V), 3% PVP (M / V), 2% β-mercaptoethanol (V / V); water bath at 65°C for 1 hour, and shake gently fr...
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