How can clone the complete miRNA primary transcript ? - (Nov/02/2007 )
There are 2 clustered miRNA. I want to know whether they are cotranscribed . Is there any good way to solve the problem?
And , If I want to clone the complete primary transcript , what should I use? I noticed that RACE mey require the RNA with polyA end.
miRNA primary transcripts are capped and poly-A tailed and can be cloned by conventional RACE (for example see Cai et al 2004, RNA 10, 1957-1966; Xie et al 2005, plant physiology 138,2145-2154). Also pre-miRNAs resulting from drosha processing can be cloned, by pcr after circularization (see Basyuk et al. 2003, NAR 31, 6593-6597)
I'm afraid I failed in my RACE expriment