Pyrosequencing is particularly suitable for SNP genotyping, since genotyping and also for this purpose, following steps are used :
(i) procure sequence information generated in the SNP identification programme,
(ii) design sequencing primers close to identified SNP sites;
(iii) amplify SNP loci using one biotinylated single strand primers;
(iv) separate biotinylated single strands by magnetic strand separation on streptavidin-coated beads,
(v) use liquid phase pyrosequencing machine, with above primers and the bead bound temple for pyrosequencing a few bases.
|
