The SSR amplified results of soybean was studied to optimize several factors applied in PCR technique system.We made L16(44)orthogonal designs for optimizing the main factors of the SSR2PCR reaction system of Soybean,and the SSR reaction was finally optimized. The results showed that there are significant effects on the results of SSR2PCR under the different levels of every factor,and the optimized SSR-PCR reaction systemwas:2.0μL 10 ×PCR Buffer,30ng DNA template,150μmol/ L dNTP,0.4μmol/ L SSR primer,1.5UTaq DNA polymerase,2. 0 mmol/L Mg2+,adding ddH2O to terminal volume 20.0 μL.The optimized PCR amplification system started at predenaturation for 5 min at 94 ℃,followed by 35 cycles of denaturation for 30 s at 94 ℃,anneal for 1min at 50 ℃,extension for 1 min at 72 ℃,the amplification was completed after extension for 5 min at 72 ℃,then stored at 4 ℃. Seventy2four primer pairs were selected from 200 primer pairs for the SSR analysis based on their reproducible and clear banding patterns,which could be used for further research.
SU Hu
,
LI Zhi-gang
,
SONG Shu-hong
. Optimization of SSR-PCR System on Soybean by Orthogonal Design and SSR Primer Selection[J]. Acta Agriculturae Boreali-Sinica, 2009
, 24(2)
: 99
-102
.
DOI: 10.7668/hbnxb.2009.02.021
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