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. 2004 Jul 7;32(12):e98. doi: 10.1093/nar/gnh094

Figure 5.

Figure 5

(A) Four and three blocks were easily amplified using five groups of 60 and 90 nt oligonucleotides separately by two-step successive PCR. M is DNA marker DL2000. Lane 1: 460 bp block; lanes 2 to 5: four 500 bp blocks; lane 6: 750 bp block; lanes 7 and 8: 800 bp blocks. (B) A full-length vip3aI gene was amplified using two-step successive PCR. M is λ-DNA Hind III marker. Lanes 1 and 2: the full-length vip3aI gene amplified from five blocks using 60 nt oligonucleotides (lane 1, annealing temperature 58°C; lane 2, 56°C); lanes 3 and 4: the full-length vip3aI gene amplified from three blocks using 90 nt oligonucleotides (lane 3, annealing temperature 58°C; lane 4 56°C). (C) Synthesis of the vip3aI gene using single-step successive PCR. M is λ-DNA Hind III marker. Lane 1: the full-length vip3aI gene amplified using 60mer oligonucleotides by single-step successive PCR; lane 2: the full-length vip3aI gene amplified using 90mer oligonucleotides by single-step successive PCR; lanes 3 and 4: the amplified vip3aI gene using 30 pmol of the two outermost primers.