松口蘑SRAP反应体系的优化

Optimization of SRAP reaction systems for Tricholoma matsutake

以松口蘑为材料,建立了松口蘑SRAP反应的优化体系。即在20μL反应体积中,模板DNA10ng,引物浓度0.3μmol/L,dNTP250mmol/L,MgCl22.5mmol/L,Taq DNA聚合酶0.5U,1×Buffer;反应程序为:94℃预变性5min,94℃变性1min,35℃退火1min,72℃延伸1min,共5个循环;94℃变性1min,50℃退火1min,72℃延伸1min,共35个循环;72℃延伸5min。

Optimized SRAP reaction systems for Tricholoma matsutake were established. Namely, in the 20μL reaction volume, template DNA was 10ng, primer concentration 0.3 μmol/L, dNTP 250mmol/L, MgCl2 2.5mmol/L, Taq DNA polymerase 0.5 U and 1×Buffer. Reaction procedttres were that： predegenerating 5 min at 94℃, degenerating 1 min at 94℃, annealing 1 min at 35℃ and extending 1 min at 72℃, and 5 cycles were included; degenerating 1 rain at 94℃, annealing 1 min at 50℃ and extending 1 rain at 72℃, and 35 cycles were included; extending 5 rain at 72℃.