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Table 1 Sequence specific oligonucleotide primers, probes and temperature profiles for Pvkelch12 and Pvpm4 genes

From: Polymorphisms in Pvkelch12 and gene amplification of Pvplasmepsin4 in Plasmodium vivax from Thailand, Lao PDR and Cambodia

Gene

Method

Fragment

Primer name

Sequence (5′–3′)

PCR condition

Annealing Temperature (°C)

MgCl2 (mM)

No. of PCR cycle

Estimated PCR product Size (bp)

Pvkelch12

PCR

Nest 1

K12_F1-F

CCATACGTAAACGCTGCAAAT

58

2

25

2139

K12_c.2216R

CTCCCCATCTGTTCCATGTC

F1

K12_F1-F

CCATACGTAAACGCTGCAAAT

58

2

30

980

K12_F1-R*

TTCTTAATTTGTTTATACCCGTTTGA

F2

K12_F2-F

CAAGCTTTTTAAAGACAAAAAGGAA

58

2

30

728

K12_F2-R*

CAGTTTCGAAAAGGGCTTTATAATC

F3

K12_c.1304F*

TGGTTTCGATGGGGTAGAGT

58

2

30

912

K12_c.2216R

CTCCCCATCTGTTCCATGTC

Pvpm4

PCR

Nest 1

PV_PM4F1_F

TCAAAAGGAGTACGAAGCATACAA

62

2

30

1703

PV_PM4F2_R

TGTTCTAATTACAGCACCAACACA

F1

PV_PM4F1_F*

TCAAAAGGAGTACGAAGCATACAA

62

2

30

807

PV_PM4F1_R*

ATGGGTTCTAAATCATCAGTGTCA

F2

PV_PM4F2_F*

GATGCAGCATTAAAAATCTGTACG

62

2

30

1086

PV_PM4F2_R

TGTTCTAATTACAGCACCAACACA

Pvpm4

Vector-based

primer

Pvpm4

CPvPM4-F*

CPvPM4-R*

AAAATCGAGAGACCCTATGACAAG

ATGGGTTCTAAATCATCAGTGTCA

62

2

35

461

Pv β-tubulin

CPvtubulin-F*

AAATTAGGGAAGAATACCCAGACC

62

2

35

451

CPvtubulin-R*

TCACTTGCACACATCATATTCTTG

Pvpm4

Real-time PCR

Pvpm4

Pvpm4_F(CNV)

Pvpm4_R(CNV)

PvPm4_Probe(CNV)

ACCACCAAAAGTTTATGCTCATC

CGTAAGACTTAGACTTGCTAGAGTCG

6FAM-CAAAAAATGCAATAGCAGCGGA-TAMRA

58

–

50

133

Pv β-tubulin

PvTubulin_F(CNV)

PvTubulin_R2(CNV)

PvTubulin_Probe(CNV)

GATAATGAGGCCTTGTATGATATTT

AGGAAATCTTAACGAACATGTAACT

HEX-TGGTTTCTGCTGCCATGTCAGG-TAMRA

58

–

50

123