![Fig. 1. Multiplex detection of wild-type and mutated FII (G20210A) and FV (G1691A) using whole blood (2 μL) as the DNA source in PCR. Lanes 1 through 3, undigested multiplex PCR products using the described conditions for FII (lane 1 [32 cycles] and 3 [40 cycles]) and for FV (lane 2). Lanes 4 through 9, undigested and Mnl I and HindIII double-digested PCR product electrophoresis of a patient double heterozygote for FII and FV; undigested and digested factor FV fragment (lanes 4 and 5, respectively); undigested and digested FII fragment (lanes 6 and 7, respectively); undigested and digested FII and FV fragments obtained after multiplex PCR (lanes 8 and 9, respectively). Lanes 10 through 13, double-digested products obtained after multiplex PCR from a normal individual (lane 10), an FII and FV Leiden double heterozygote (lane 11), an FII heterozygote (lane 12), and an FV Leiden heterozygote (lane 13). Products were separated on a 2.5% agarose gel (per lane, 20 μL of the digestion mix is loaded). Marker is a 100-bp size marker (GIBCO BRL, Grand Island, NY). Relevant fragment sizes in basepairs are indicated.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/91/6/10.1182_blood.v91.6.2208/2/blod4044201.jpeg?Expires=1725323810&Signature=sz0Q3IRCb92IQOyYAIe5k2LMFR8ivRlbG1hiE2jUz-GVUVyZzKeHSDxtuUmDDR6tQ6pz5VdqZ0gPs05GJ6uf5LO1cU9-BWzcSzX95uQMePvkczYjrBmI4HxAa1r7zzAhf7J9sctdW1n-NiCpc94wt1zOj84T~GbyDwNoX~cyN4~iaM2kBaTMiXGWx4FHXbe5tLhpFZzDLQyFcluJ1eSbjYkPlT~DLtIzKRMh0htM0BYkWlJUA~BKm8oG11HjS-cLu9nKpJqU347LAWpx43P1lc1d5EESsRU4FkHQTTZeaMAgUtIzw5FOn~qSkRGmXO~URNA3SrzXKyest2QA-9ivXA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Multiplex detection of wild-type and mutated FII (G20210A) and FV (G1691A) using whole blood (2 μL) as the DNA source in PCR. Lanes 1 through 3, undigested multiplex PCR products using the described conditions for FII (lane 1 [32 cycles] and 3 [40 cycles]) and for FV (lane 2). Lanes 4 through 9, undigested and Mnl I and HindIII double-digested PCR product electrophoresis of a patient double heterozygote for FII and FV; undigested and digested factor FV fragment (lanes 4 and 5, respectively); undigested and digested FII fragment (lanes 6 and 7, respectively); undigested and digested FII and FV fragments obtained after multiplex PCR (lanes 8 and 9, respectively). Lanes 10 through 13, double-digested products obtained after multiplex PCR from a normal individual (lane 10), an FII and FV Leiden double heterozygote (lane 11), an FII heterozygote (lane 12), and an FV Leiden heterozygote (lane 13). Products were separated on a 2.5% agarose gel (per lane, 20 μL of the digestion mix is loaded). Marker is a 100-bp size marker (GIBCO BRL, Grand Island, NY). Relevant fragment sizes in basepairs are indicated.
