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Table 1 Pfdhfr , pfdhps , pfmdr1 and pfcrt PCR primers sequences used in amplification reactions

From: Prevalence of molecular markers of drug resistance in an area of seasonal malaria chemoprevention in children in Senegal

Genes names

Primers names

Primers sequences

amplicon size (bp)

PCR cycling conditions

Pfdhfr

PCR1

dhfr_M1

5′-TTTATGATGGAACAAGTCTGC-3′

650

93°C for 5min/(94°C for 30s- 54°C for 60s- 65°C for 60s) × 41cycles/ 65° for 5min/ 15°C for 5min

Nested

dhfr_M7

5′-CTAGTATATACATCGCTAACA-3′

  
 

dhfr-M9

5′-CTGGAAAAAATACATCACATTCATATG-3′

594

95°C for 5min/(93°C for 30s- 56°C for 30s- 68°C for 75s) × 30 cycles/ 75°C for 5min

Sequencing

dhfr-M3

5′-TGATGGAACAAGTCTGCGACGTT-3′

  
 

dhfr-M9

5′-CTGGAAAAAATACATCACATTCATATG-3′

 

96°C for 1min/(96°C for 30s- 50°C for 30s- 60°C for 4min) × 26 cycles/4°C hold until ready to purify

 

dhfr-M3

5′-TGATGGAACAAGTCTGCGACGTT-3′

  

Pfdhps

PCR1

dhps-N1

5′-GATTCTTTTTCAGATGGAGG-3′

770

94°C for 3min/(94°C for 30s- 55°C for 30s- 65°C for 60s) × 30cycles / 65°C for 5min/ 15°C for 5min.

 

dhps-N2

5′-TTCCTCATGTAATTCATCTGA-3′

711

 

Nested

dhps-R2

5′-AACCTAAACGTGCTGTTCAA-3′

 

94°C for 5min/(94°C for 30s- 60°C for 30s- 65°C for 1min) × 30cycles /65°C for 5min/15°C for 5min.

 

dhps-R/

5′-AATTGTGTGATTTGTCCACAA-3′

  
 

dhps-R2

5′-AACCTAAACGTGCTGTTCAA-3′

 

96°C for 1min/(96°C for 30s-50°C-60°C for 4min) × 26 cycles/ 4°C hold until ready to purify.

Sequencing

dhps-R/

5′-AATTGTGTGATTTGTCCACAA-3′

  

Pfmdr1

PCR1

fn1/1

5′-ACAAAAAGAGTACCGCTGAAT-3′

578

94°C for 3min/(94°C for 30s- 55°C for 30s- 65°C for 1min) × 30cycles/ 65°C for 5min/ 15°C for 5min.

 

rev/c1

5′-AAACGCAAGTAATACATAAAGTC-3′

534

 

Nested

mdr2/1

5′-ACAAAAAGAGTACCGCTGAAT-3′

 

94°C for 3min/(94°C for 30s- 60°C for 30s- 65°C for 1min) × 30cycles/ 65°C for 5min/ 15°C for 5min.

 

newrev1

5′-AAACGCAAGTAATACATAAAGTC-3′

  

Sequencing

mdrfr1f-seq

5′-GTCGAATTATTTAGAAAAAT-3′

 

96°C for 1min/(96°C for 30s- 50°C -60°C for 4min) × 26cycles/4°C hold until ready to purify.

 

mdrfr1r-seq

5′-GCAAGTAATACATAAAGT-3′

  

Pfcrt

 

Crtd1

5′-TGTGCTCATGTGTTTAAACTT 5′-

166

94°C for 3min/(94°C for 30s- 55°C for 30s- 65°C for 1min) × 30 cycles/ 65°C for 5min/15°C for 5min.

 

Crtd2

CAAAACTATAGTTACCAATTTTG

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