Hi,
Attempted to extract cfDNA from a 4 mL plasma sample, with poor yield. Went through the manual in the section regarding poor yield, and feel that this does not apply. Some potential issues:
- using water rather than Buffer Ave to elute-->does this affect yield?
- QIAVac using lab wall vacuum-- we also bought a vacuum regulator, which only reaches pressure of -800 mbar. Could this be an issue?
- is it possible to use centrifuge for buffers/ethanol washes rather than vacuum for better yield?
TIA.
Hello,
using water instead of Buffer AVE can affect yield negatively. Switching to water might reduce the yield and integrity of the cfDNA.
A vacuum pressure of -800 mbar might be insufficient. The QIAvac system typically requires a higher vacuum pressure.
Centrifugation can sometimes provide more consistent results when used for ethanol washes.
Good luck ;-)
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