I extracted RNA from two different types of cell lines: one that is sensitive to cisplatin and one that is resistant. I then followed the cDNA synthesis protocol and observed a strong band for the resistant cell line cDNA, but no band for the sensitive cell line. Can you help me identify the problem?
Hi Zahra Firoozi
Possible reasons include: low RNA quality or concentration, the presence of inhibitors or contaminants, inappropriate priming strategy, issues with the reverse transcriptase enzyme, suboptimal reaction conditions, or RNA molecules' integrity and size.
So you should check the RNA quality and concentration, consider purification steps if needed, verify the priming strategy, and ensure proper storage and functionality of the reverse transcriptase. Optimization of reaction conditions may also be necessary. Running positive control reactions can also help pinpoint the problem.
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