Abstract
A novel single-capillary method of verification of both-strand analysis of the -1552delAGG genetic polymorphism in the TGFB1 gene is presented. For the verification we used the single-tube sequencing reaction with forward and reverse primers modified at the 5'-end by adapters composed of 55 (forward primer) and 10 (reverse primer) repetitive units of cytosine and adenine (CA adapters).
The CA adapters made it possible to get complete readings (of both strands of the amplicons) and to determine different migration times for forward and reverse sequencing fragments in single-capillary electrophoresis. Our study showed that the attachment of the CA adapters to the primers made it possible to perform sequencing of both strands in one PCR tube as well as to separate forward and reverse sequencing fragments in one capillary.