Researchers can now source stable cDNA (complementary DNA) from ECACC for pre-screening transcriptomes for genomic expression characteristics.
Our ECACC scientists use high quality RNA (with RIN scores typically above 9.7) to generate cDNA from ECACC’s diverse range of human cell lines. We use PCR with primers designed to intronic sequences of housekeeping genes to confirm the absence of genomic DNA and RT-PCR to detect human cDNA sequences that do not co-amplify processed pseudogenes* to confirm the presence of cDNA in the final product.
cDNA is available from all the human cell lines in the following collections – please search or browse for the cell line of interest by name or catalogue number:
ECACC scientists generated cDNA from cell line 1321N1 to test the principle of using cell line cDNA as a transcriptome pre-screen to test for the presence of interesting biomarkers. We chose 1231N1 because it is a well characterised cell line and the presence of a class of G-protein coupled receptor, the muscarinic sub-type three receptor, is well-established in a range of pharmacological analyses. It was fairly simple for us to identify a PCR product representing the muscarinic sub-type three receptor using cDNA from this cell line and in addition, we found a second PCR product that represented the muscarinic sub-type five receptor which was previously unreported in this cell line.
* Raff, T., et al. Design and testing of β-actin primers for RT-PCR which do not co-amplify processed pseudogenes. 1997. Biotechniques. 23:456-460
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