RNA which has been extracted from blood collected in Heparin tubes are not showing any amplification after cDNA synthesis, in comparison to Blood of same patient collected in ETDA tubes. Any suggestions?
Hey Vera,
Are you looking at cfRNA? What kit are you using to isolate the RNA?
Heparin is known to inhibit reverse transcriptase activity (Li et al., 2002) so you may want to use a blood collection tube that won't. Have you tried using different preservatives? If not, I'd recommend trying out Norgen's Blood Preservation Tubes. They consistently perform better than EDTA tubes and even some of the more popular brands out there, in terms of yield and limiting gDNA contamination.
Li, C. M., Newman, D., Khosla, J., & Sannes, P. L. (2002). Heparin inhibits DNA synthesis and gene expression in alveolar type II cells. American journal of respiratory cell and molecular biology, 27(3), 345-352.
Norgen's Preservative Tubes
https://norgenbiotek.com/product/cf-dnacf-rna-preservative-tubes
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