I am working with Pyromark Q24 protocol on bisulfite converted DNA that has been through a PCR and verified by gel. I have a very high substrate peak (~200-250) that is resulting in low nucleotide peaks in my pyrograph. Qiagen's FAQ attributes this to pyrophosphate or dATP contamination. Can this be fixed by optimizing the PCR cycles or increasing the amount of DNA into the bisulfite reaction? Any suggestions are helpful in troubleshooting this issue.
I am working with Pyromark Q24 protocol on bisulfite converted DNA that has been through a PCR and verified by gel. I have a very high substrate peak (~200-250) that is resulting in low nucleotide peaks in my pyrograph. Qiagen's FAQ attributes this to pyrophosphate or dATP contamination. Can this be fixed by optimizing the PCR cycles or increasing the amount of DNA into the bisulfite reaction? How do I lower my substrate peak?
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