SNaPshot® genotyping is a primer extension-based technique that enables multiplexing of several single nucleotide polymorphisms (SNPs) at known locations in one single tube.
The chemistry behind this technology is based on the dideoxy single-base extension of unlabeled primer; the primer binds to the complementary template in the presence of fluorescently labelled ddNTPs and AmpliTaq DNA polymerase. The polymerase extends the primer by 1 nucleotide, adding a single ddNTP to its 3’ end.
Run Fragment Analysis on ABI Sequencer and analyze the results using GeneMapper v4.0 software.
|wt/mutant||31C(black) and 31A(green)||26C(black) and 26T(red)|
In one genotyping result, you will see both CFTRexon5 and CFTRexon11 peaks.