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Essay Details:
Essay text:
In the case of thiopurine
S-methyltransferase (TPMT), differentiating among
TPMT*3A, *3B, and *3C alleles is sometimes necessary
for predictive genotyping.
Methods: The genomic region including the two SNPs
that define TPMT*3A, *3B, and *3C alleles was amplified
by long-range PCR. The resulting PCR product was
circularized by ligation and haplotyped by allele-specific
amplification PCR followed by product identification
with hybridization probes.
Results: Critical points were the long-range PCR conditions,
including choice of buffer and primers, optimization
of the ligation reaction, and selection of primers
that allowed for strict allele-specific amplification in the
second-round PCR...
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