Poly(A) polymerase modification and reverse transcriptase PCR amplification of environmental RNA

A protocol that couples poly(A) polymerase with a multifunctional reverse transcriptase (RT) for targeted cloning of environmental prokaryotic cDNA is described. The reverse transcriptase PCR (RT-PCR) cDNA synthesis kit demonstrates the feasibility of directly accessing prokaryote RNA (rRNA and/or mRNA) in environmental samples, but the results also show potentially important problems.