5端RACE升级版
互联网
完成这个RACE需要全套反转录系统,当然选一个好的反转录酶(无RNase H),dUTP,taq酶,Uracil DNA glycosylase ,还有这几条引物:
Adapter A AUCUCGAGUUCGCGCCGGAUCC(T) 25 VN cDNA synthesis and adapter-A incorporation
Adapter B AUAUGCACUGCCGCGUCUGAGGGGGGGG Cap finder adapter-B incorporation
Primer A CUCGAGUUCGCGCCGGAUC Second-strand cDNA synthesis
Primer B AUAUGCACUGCCGCGUCUGA Second-strand cDNA synthesis
5 T-RACE primer ATATGCACTGCCGCGTCTGA 5 specific amplification of cDNA ends 3 T-RACE primer CTCGAGTTCGCGCCGGATC 3 specific amplification of cDNA ends
实验原理
原理图: