黑木相思双低频酶cDNA-AFLP反应体系的建立

Establishment of cDNA-AFLP Analysis System with Two Low-frequency Restriction Enzyme in Acacia melanoxylon

以黑木相思组培苗为实验材料,对影响cDNA-AFLP反应体系的关键因素进行了研究,建立了适用于黑木相思的cDNA-AFLP分析体系.结果表明:适用于黑木相思dscDNA的限制性酶切组合为EcoRⅠ和PstⅠ,dscDNA酶切用量为500 ng.用作预扩增模板的连接产物稀释倍数为1倍,用作选择性扩增模板的预扩增产物稀释倍数为50倍.

An optimized system was established after studying on the factors affecting cDNA- AFLP analysis system of Acacia melanoxylon cultured in vitro. The results indicated that the restric- tion enzyme combination was EcoR I and Pst I , the dosage of dscDNA was 500 ng, the ligation products using as template of pre-amplification were diluted to one time, and the pre-amplification products using as template of select-amplification were diluted to 50 times.