I have tried all protocols and I can´t avoid degradation so I don't get any amplicon in the PCR reaction. I tried trizol, regular extraction from wormbook, different lysis buffer, etc. Does someone have any protocol or advice for both steps?
Thank you!
Katie A S Burnette
Store your DNA extractions in the freezer. Ask to borrow some good quality DNA to use as a positive control in your PCR reactions. The issue might be with the PCR, not with your DNA.
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Maxime Pichon
Do you freeze all your extract before using PCR? If your process was always extraction>freezing>PCR, I would recommend to apply PCR just after the extraction, assuring an optimized concentration.
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Eliana Fernandez
The positive control in the PCR was OK, and I tried extracting and PCR right after. Thank you anyway!
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