4. Principle
Gap-PCR
• PCR primer pairs are designed to flank a known deletion
(Faa et al., 1992).
• When the deletion is vast (>2 kb) it is technically difficult to
generate a product.
• As a control, a primer can be included that anneals within the
deleted sequence to generate an additional product with one
of the other primers flanking the deletion. In this way, false
negatives are monitored in the wild type and indicates if an
individual is heterozygous for the deletion.
5. Principle
Primer are
1. A4
2. A1B
3. A9
Product are
Normal allele : A4/A1B (194bp)
Mutant allele : A4/A9 (550 bp)
6. Applications
• Gap-PCR is also routinely used to detect a
deletion gene. Such as in Thallasemia.
• Can also apply for another deletion mutation.
