High Quality 454-Ready cDNA is made from our custom or catalog cDNA libraries. Briefly, library DNA is digested with Not I and in vitro RNA transcripts are produced using the SP6 RNA polymerase promoter. Then, first strand cDNA is made from these transcripts using a modified primer adapter that reduces the size of the poly A sequence (to about 20 As). After the second strand is synthesized, the double stranded (ds) cDNA is blunt ended and size fractionated. This ds cDNA is now ready to be used with the 454 sequencing method or with other next generation sequencing technology (i.e., illumina). We guarantee at least 6 µg of ds cDNA, 150 ng/µl concentration in TE and 1 kb average cDNA size, however, we generally produce 8 µg of cDNA, 200 ng/µl concentration and 1.5 kb average cDNA size.
We will make 454-ready cDNA from our custom or catalog libraries or directly from customer supplied cDNA libraries, tissue or RNA.
Quality testing: Gel analysis of cDNA population.
Shipment and storage conditions: 454-ready cDNA will be shipped in dry ice. Storage of cDNA at -80°C is recommended.