Archives of Medicine

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Improved probes for detection of V617F mutation in chromosome 9-borne JAK 2 gene linked to conditions of Polycythemia Vera and other myeloproliferative disorders

Pratap N Mukhopadhyaya

Nucleotide positions 5063534 to 5063554 and 5063973 to 5063952 within the Human Janus Kinase 2 (JAK2) gene (Gene bank accession number NT_008413.17) were identified as novel, optimized oligonucleotide primer hybridization sites for thermal amplification of a robust 440 bp PCR amplicon which encompassed the V617F mutation linked to occurrence of Polycythemia vera (PV), a well known myeloproliferative disorder (MPD) in humans. The single strand oligonucleotide primer pair was also found suitable to generate high quality fluorescent nucleotide sequence data from both strands of DNA that was sufficient for detection of all possible genotypes of JAK2 gene with regard to V617F mutation. The accuracy and reproducibility of the method was satisfactory ((r2=0.99, p