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Table 1 pfmdr1 PCR primer sequences and reaction conditions used in Fragments 3 and 4 amplification reactions

From: Markers of anti-malarial drug resistance in Plasmodium falciparum isolates from Swaziland: identification of pfmdr1-86F in natural parasite isolates

Gene Fragment

Primer name

 

Primer Sequence

Codons

PCR Cycling conditions

Fragment 3

Primary FR3

MDRF3N1

F

5'-GCATTTTATAATATGCATACTG-3'

1034, 1042

94°C 3 min/[94°C 30 s-56°C 60 s 65°C 50 s]

× 30 cycles

65°C 5 min/15°C 5 min

 

MDRF3R1

R

5'-GGATTTCATAAAGTCATCAAC-3'

  

Nested FR3

MDRF3N2

F

5'-GGTTTAGAAGATTATTTCTGTA-3'

  
 

MDRF3R1

R

5'-GCATTTTATAATATGCATACTG-3'

  

Fragment 4

Primary FR4

MDRF4N1

F

5'-CAAACCAATCTGGATCTGCAG-3'

1246

94°C 3 min/[94°C 30 s-55°C 60 s-65°C 40 s]

× 30 cycles

65°C 5 min/15°C 5 min

 

MDRF4R1

R

5'-CAATGTTGCATCTTCTCTTCC-3'

  

Nested FR4

MDRF4N2

F

5'-GATCTGCAGAAGATTATACTG-3'

  
 

MDRF4R1

R

5'-CAATGTTGCATCTTCTCTTCC-3'

  

Sequencing

FR3

MDRF3N2

F

5'-GGTTTAGAAGATTATTTCTGTA-3'

 

96°C 1 min

[96°C 30 s-50°C-60°C 4 min]

× 26 cycles

 

MDRF3R1

R

5'-GCATTTTATAATATGCATACTG-3'

  

FR4

MDRF4N2

F

5'-GATCTGCAGAAGATTATACTG-3'

  
 

MDRF4R1

R

5'-CAATGTTGCATCTTCTCTTCC-3'

  
  1. FR - Fragment F - forward R - Reverse
  2. NB: Cycling conditions are the same for primary and nested PCRs.