The present invention includes a method and kit for cDNA synthesis of a 3′UTR/poly(A) tail junction of cellular RNA comprising: obtaining RNA comprising a 3′UTR/poly(A) junction and a poly(a) tail; combining the RNA with three terminating nucleotides of modified-deoxyGTP, modified-deoxy...
A tight band of DNA from 130–150 bp was isolated with a clean razor blade and purified using the MinElute Gel Extraction Kit (Qiagen). The Quick Blunting Kit (NEB) was used to blunt the ends of the DNA in a 25 μl reaction volume containing 2.5 μl 10× Blunting Buffer, 2.5 μl...
The rRNAs were depleted by using Epicentre Ribo-Zero kit. The RNAs were ligated to biotinylated 3′ adaptor, partially digested by RNase T1. The fragmented RNAs were pulled down, phosphorylated, and gel purified (500–1000 nt). The purified RNAs were ligated to 5′ adaptor, reverse ...