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Table 1 Sequences of the primers used to amplify the Pvmsp-3α and Pvmsp-3β genes of Plasmodium vivax and the Pfmsp-1 and Pfmsp-2 genes of Plasmodium falciparum isolates from Bannu district, Pakistan.

From: Genetic structure of Plasmodium vivax and Plasmodium falciparum in the Bannu district of Pakistan

Gene (PCR reaction)§ Primers* PCR cycling conditions** Reference
P. vivax    
Pvmsp-3α (N1) F: 5'-CAGCAGACACCATTTAAGG-3' 95°C 3 min/[94°C 30 s, 54°C 30 s, 68°C 2.5 min] × 35 cycles, 68°C 5 min [19]
  R: 5'-CCGTTTGTTGATTAGTTGC-3'   
Pvmsp-3α (N2) F: 5'-GACCAGTGTGATACCATTAACC-3' 95°C 3 min/[94°C 30 s, 55°C 30 s, 68°C 2.5 min] × 30 cycles, 68°C 5 min [19]
  R: 5'-ATACTGGTTCTTCGTCTTCAGG-3'   
Pvmsp-3β (N1) F: 5'-GTATTCTTCGCAACACTC-3' 95°C 3 min/[94°C 30 s, 54°C 30 s, 68°C 2.5 min] × 35 cycles, 68°C 5 min [20]
  R: 5'-CTTCTGATGTTATTTCCAG-3'   
Pvmsp-3β (N2) F: 5'-CGAGGGGCGAAATTGTAAACC-3' 95°C 3 min/[94°C 30 s, 55°C 30 s, 68°C 2.5 min] × 30 cycles, 68°C 5 min [20]
  R: 5'-GCTGCTTCTTTTGCAAAGG-3'   
P. falciparum    
Pfmsp-1 (N1) F: 5'-CTAGAAGCTTTAGAAGATGCAGTATTG-3' 95°C 5 min/[94°C 30 s, 55°C 30 s, 72°C 1 min] × 35 cycles, 72°C 5 min [17]
  R: 5'-CTTAAATAGTATTCTAATTCAAGTGGATCA-3'   
K1 family (N2) F: 5'-AAATGAAGAAGAAATTACTACAAAAGGTGC-3' 95°C 5 min/[94°C 30 s, 56°C 30 s, 72°C 45 s] × 30 cycles, 72°C 5 min [17]
  R: 5'-GCTTGCATCAGCTGGAGGGCTTGCACCAGA-3'   
MAD20 family (N2) F: 5 '-AAATGAAGGAACAAGTGGAACAGCTGTTAC-3 ' 95°C 5 min/[94°C 30 s, 56°C 30 s, 72°C 45 s] × 30 cycles, 72°C 5 min [17]
  R: 5 '-ATCTGAAGGATTTGTACGTCTTGAATTACC-3 '   
RO33 family (N2) F: 5 '-TAAAGGATGGAGCAAATACTCAAGTTGTTG-3 ' 95°C 5 min/[94°C 30 s, 56°C 30 s, 72°C 45 s] × 30 cycles, 72°C 5 min [17]
  R: 5 '-CATCTGAAGGATTTGCAGCACCTGGAGATC-3 '   
Pfmsp-2 (N1) F: 5 '-ATGAAGGTAATTAAAACATTGTCTATTATA-3 ' 95°C 5 min/[94°C 30 s, 56°C 30 s, 72°C 45 s] × 30 cycles, 72°C 5 min [17]
  R: 5'-CTTTGTTACCATCGGTACATTCTT-3'   
FC27 family (N2) F: 5'-AATACTAAGAGTGTAGGTGCARATGCTCCA-3' 95°C 5 min/[94°C 30 s, 56°C 30 s, 72°C 45 s] × 30 cycles, 72°C 5 min [17]
  R: 5 '-TTTTATTTGGTGCATTGCCAGAACTTGAAC-3 '   
3D7/IC family (N2) F: 5 '-AGAAGTATGGCAGAAAGTAAKCCTYCTACT-3 ' 95°C 5 min/[94°C 30 s, 56°C 30 s, 72°C 45 s] × 30 cycles, 72°C 5 min [17]
  R: 5 '-GATTGTAATTCGGGGGATTCAGTTTGTTCG-3 '   
  1. § N1 = Nest 1 (Primary) reaction; N2 = Nest 2 (Secondary) PCR reaction
  2. * F= Forward primer; R = Reverse primer. The reference sources of the primers are indicated.
  3. **The cycling conditions have been modified in the present work.