A multiplex PCR for detection of knockdown resistance mutations, V1016G and F1534C, in pyrethroid-resistant Aedes aegypti

Table 1 Sequences of primers used in this study

Primer name	Primer sequence (5′-3′)	Product size (bp)	Exon^a
Direct sequencing
IIP_F	GGTGGAACTTCACCGACTTC	581	15
IIS6_R	GGACGCAATCTGGCTTGTTA	581	16
Ge-IIIS6_F	GCTGTCGCACGAGATCATT	635	23
IIIS6_R	GTTGAACCCGATGAACAACA	635	25
Multiplex PCR
1016 genotyping
Gly1016f	ACCGACAAATTGTTTCCC		15–16^b
Val1016r	[short GC tail]^c AGCAAGGCTAAGAAAAGGTTAATTA	60	16
Gly1016r	[long GC tail]^d AGCAAGGCTAAGAAAAGGTTAACTC	80	16
1534 genotyping
c1534-f	GCGTACCTGTGTCTGTTCCA	368	23
c1534-r	GGCTTCTTCGAGCCCATCTT	368	24
Ae1534F-r	GCGTGAAGAACGACCCGA	232	24
Ae1534C-f	CCTCTACTTTGTGTTCTTCATCATCTG	180	24

^aExon from the Ae. aegypti VGSC gene. This transcript corresponds to VectorBase Transcript ID AAEL006019
^bIntron between exon 15 and 16
^cShort GC tail sequence: 5′-GCG GGC-3’
^dLong GC tail sequence: 5′-GCG GGC AGG GCG GCG GGG GCG GGG CC-3′

ISSN: 1756-3305