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SNP Melt Curves
U937 Leukemia Single Cells were amplified using the GenomePlex® Single Cell Whole Genome Amplification procedure. Post reaction products were removed by the GenElute™ PCR Clean-Up Kit (Product Code NA1020). The single cell DNA was quantified by nanodrop and 10 nanograms of DNA was then utilized in a MGB Eclipse™ SNP quantitative PCR reaction on the ABI 7700. The MGB Eclipse™ Probe System facilitates allelic discrimination due to its ability to allow post-PCR dissociation curves. Dissociation curves allow determination of the melting point (Tm). Different alleles will have a substantially different Tm value. FAM and TET fluorescent signals help distinguish homozygous wild, heterozygous and homozygous mutant types. The WGA amplified single cell DNA was tested for 3 SNPs, MTHFR01, IGF1R-03 and NAT1-01. A positive unamplified control was evaluated with each group. The data indicates that all of the single cells were the same allele for each SNP tested.
MTHFR01
MTHFR01 Single Cell WGA Samples - FAM
MTHFR01 Control DNA Samples - FAM
MTHFR01 Control DNA Samples - TET
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IGF1R-03
IGF1R-03 Single Cell WGA Samples - FAM
IGF1R-03 Control DNA Samples - FAM
IGF1R-03 Control DNA Samples - TET
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NAT1-01
NAT1-01 Single Cell WGA Samples - TET
NAT1-01 Control DNA Samples - FAM
NAT1-01 Control DNA Samples - TET
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