2.6. Construction of the mutant DNA fragments
Synthetic mutant DNA fragments were constructed for Hb
Heraklion, Hb Adana, Hb Questembert (in α1) and Hb Questembert
(in α2) variants due to the lack of representative clinical samples. All
mutant DNA fragments were created by a three-step PCR (PCR-A,
PCR-B and PCR-AB) using genomic DNA as template as previously
described [12]. The amplified mutant DNA fragments were cloned into
the pDrive Cloning Vector using PCR cloning kit from Qiagen according
to the manufacturer's instructions.