Long cDNA Synthesis
The iScript Select cDNA synthesis kit facilitates synthesis of cDNA longer than 6 kb. First-strand cDNA was produced from 1 µg total RNA using the iScript Select cDNA synthesis kit and the provided oligo(dT) primer mix. A 2 µl aliquot of the 20 µl cDNA synthesis reaction was subjected to 35 PCR cycles using a proofreading high-fidelity polymerase and human adenomatous polyposis coli (APC) primer sets. From a 50 µl PCR reaction, 10 µl was analyzed on a 1% agarose gel. Lane 1, 1 kb ladder; lanes 2 and 3, 6.4 kb APC PCR product; lanes 4 and 5, 8.2 kb APC PCR product.