SNaPshot® Genotyping

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.

 
Fluorescence
 Genotypes CFTRexon5 CFTRexon11
wt/wt 31C(black) 26C(black)
wt/mutant 31C(black) and 31A(green) 26C(black) and 26T(red)
mutant/mutant 31A(green) 26T(red)

In one genotyping result, you will see both CFTRexon5 and CFTRexon11 peaks.