Fig. 10.1.

Overview and analysis of the TC RNA amplification method. (A) A TC primer (containing a bacteriophage promoter sequence for sense orientation) and a poly d(T) primer are added to the mRNA population to be amplified. First-strand synthesis occurs as an mRNA-cDNA hybrid is formed after reverse transcription and terminal continuation of the oligonucleotide primers. Following RNase H digestion to remove the original mRNA template strand, second-strand synthesis is performed using Taq polymerase. The resultant double-stranded product is utilized as template for IVT, yielding high-fidelity, linear RNA amplification of sense orientation (rippled lines). (B) Schematic similar to A, illustrating the TC RNA amplification procedure amplifying RNA in the antisense orientation (rippled lines). Adapted from Ginsberg 2005 [26]