Hey all,
I am about to do a research the requires investigating a SNP in the promoter of the bradykinin 1 receptor B1R (G-699 to C) substitution.
The paper I found uses SSCP and [32P] - alpha dCTP. Can I skip the denaturation of the PCR product using formamide dye and use normal dNTPs and EtBr for staining after digestion?
Since it is RFLP then one snp base creates a restriction site and can be determined by cutting the pcr product and either getting 2 fragments ( cuts) or 3 fragments (one allele cuts and the other does not cut) or 1 band ( neither allele cuts).
If your snp does not create a cut site you can use a mismatched primer to create a cut site from almost any sequence using the DCAPS technique to prove that any sequence exists at a particular position but labelled ntps are not necessary to show the existence of a base at a snp position
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