Myeloid Leukemia

256 Campbell et al.
Fig. 1. Allele-specific polymerase chain reaction (PCR) and BsaXI digestion methods
for detecting the JAK2 V617F mutation. (A) Allele-specific PCR generates a 364bp
product from wild-type sequence, as a result of amplification from primers F1 and
R1. V617F DNA gives a 364-bp product from amplification between F1 and R1 primers,
and a 203-bp product as a result of amplification between F2 and R1 primers. (B)
After amplification with primers F3 and R3, BsaX1 only digests the wild-type
sequence (giving 30-, 174-, and 206-bp products), but not the V617F sequence (leaving
an undigested 410-bp product). The 174- and 206-bp product co-migrate on a conventional
2% agarose gel.