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Multiplex PCR
Before proceeding to multiplex PCR, cells were lysed by
incubation at 65°C for 10 min. The alkaline lysis buffer was
then neutralized by the addition of 5 ml of neutralization
buffer.
A nested multiplex PCR assay is used to co-amplify all the
selected loci.
The first round PCR containes the external primers for the
amplification of the informative HLA STR markers selected
during the preclinical work-up of each PGD case, the gene
regions involved by mutations, STR markers linked to these
regions for ADO detection and STR markers used for detection
of aneuploidies in patients of advanced reproductive age.
The first round multiplex PCR is followed by separate second
round PCR reactions for each locus, using 2 ml of the
primary PCR products.
Next :
The HLA complex
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