A Multiplex Snapback Primer System for the Enrichment and Detection of JAK2 V617F and MPL W515L/K Mutations in Philadelphia-Negative Myeloproliferative Neoplasms
Figure 5
TaqMan probe qPCR assay for JAK2 V617F and MPL W515L/K mutation. (a) TaqMan system for JAK2 V617F detection. A sigmoidal shaped amplification curve in the fluorescent signal versus cycle number plot of FAM fluorescence channel (465 nm–510 nm) indicated the presence of JAK2 V617F mutant allele in the DNA sample, while the amplification curve in VIC channel (540 nm–580 nm) represents the wild-type JAK2 copies in the sample. JAK2 V617F and JAK2 wild-type homozygotes specifically produced the amplification curve in the FAM and VIC channel, respectively. The amplification of mutation heterozygote could be observed both in the FAM and VIC channel. (b) TaqMan system for MPL W515L/K mutations. The system was consisted of three allele-specific channels. The amplification curve of homozygous mutant and wild-type samples was only generated in the corresponding channel. The amplification curves of heterozygote with MPL W515L or MPL W515K mutation would be observed in both the wild-type channel and the mutant allele channel.
