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. 2014 Sep 24;70(1):116–123. doi: 10.1093/jac/dku358

Table 1.

Primer sequences and PCR cycling conditions for amplifying and sequencing full-length pfcrt

Gene PCR primer sequence 5′ to 3′ Cycling conditions Product size (bp)
pfcrt fragment 1 (exons 1–4) outer PCR 94°C3 min, (94°C1 min; 45°C1 min, 65°C1 min) × 30 cycles, 65°C10 min 873
CRTP1: CCG TTA ATA ATA AAT ACA CGC AG
NG1_REV: GGA TAC CAT AGC ATT TAA TC
nested PCR 94°C3 min, (94°C1 min; 50°C1 min, 65°C1 min) × 30 cycles, 65°C10 min 827
CRTP1: CCG TTA ATA ATA AAT ACA CGC AG
NG4_REV: AGC ATT TAA TCT TAA AAT GTC
pfcrt fragment 2 (exons 4–13) outer PCR 94°C3 min, (94°C1 min; 50°C1 min, 65°C1 min) × 30 cycles, 65°C10 min 874
NG5_FOR: CTC GGA GCA GTT ATT ATT G
NG3_Rev: ATT CCT TAT AAA GTG TAA TGC GA
nested PCR 94°C3 min, (94°C1 min; 50°C1 min, 65°C1 min) × 30 cycles, 65°C10 min 827
NG2_FOR: GCT CTT GTA GAA ATG AAA TTA TC
NG3_REV: ATT CCT TAT AAA GTG TAA TGC GA
5′ and 3′ UTR sequencing primers NG6_REV: GAA CAT AAT CAT ACA AAT A
NG7_FOR: CCG TGT AGG AAA TAT TAT CT
Sequencing primers for confirming intron skipping P11: TCCTTTTTCCAATTGTTCACTTC
P13: GACGGAGCATGGGTAAGAAG
P17: CAAATGGCTTGTTCGTTCATAA