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Table 3 The information of primers, including VectorBase ID, gene name, primer use and sequences. Primers were used for PCR amplification of dsRNA template DNA, in vitro transcription of dsRNA from cDNA and silencing validation by real-time quantitative reverse transcription PCR (qRT-PCR) for RNAi validation of candidate P450 genes involved in malathion resistance of Culex quinquefasciatus. Each primer set includes a forward sequence (F) and a reverse sequence (R). An additional probe (P) sequence was used for probe-based qRT-PCR.

From: Potential key genes involved in metabolic resistance to malathion in the southern house mosquito, Culex quinquefasciatus, and functional validation of CYP325BC1 and CYP9M12 as candidate genes using RNA interference

VectorBase ID

Gene Name

Primer Use

Primer sequences (5′–3′)

CPIJ014220

cytochrome P450 CYP9M12

amplification

F: CCTTTTTCTTCGGAGGTATCG

   

R: AAAAGCCGTTCTTCGCACT

  

dsRNA from cDNA

F: TAATACGACTCACTATAGGGCCATGTTGTGCTTTGCAATC

   

R: TAATACGACTCACTATAGGGGTTCCGAAATCTCCACCGTA

  

qRT-PCR

F: GTTACTGTGAGGGGGCAATG

   

P: AATCGGCTGAGGGGAGGATC

   

R: CATGCCAAACGAGATTGATG

CPIJ015958

cytochrome P450 CYP325BC1

amplification

F: GGATATGATCTGCGCAACTG

   

R: CGGCAAATATGATTTCATCCA

  

dsRNA from cDNA

F: TAATACGACTCACTATAGGGGGGAGTGGACATAAACGTGC

   

R: TAATACGACTCACTATAGGGAATGTTCCGCTATGGATTCG

  

qRT-PCR

F: AAGTCGGAGGACTGTCTGGA

   

P: TCATCCTGGAAAGCGGTTCG

   

R: GCTTGGAGACTTGCTCAACC

CPIJ000162

60 S ribosomal protein L8

qRT-PCR

F: ACTTCCGTGACCCGTACAAG

   

P: AGCTCCGCAAGCAGCTGTTC

   

R: ACCGGTCTTTTCCTCCAGAT

 

DsRed1

amplification

F: CGATGGTGTAGTCCTCGTTG

  

R: GCTCCTCCAAGAACGTCATC

 

dsRNA from cDNA

F: TAATACGACTCACTATAGGGGGTGTAGTCCTCGTTGTGG

  

R: TAATACGACTCACTATAGGGGAAGCTGAAGGTGACCAAGG

 

qRT-PCR

F: TAGTCCTCGTTGTGGGAGGT

  

P: ACGGGCTTCTTGGCCATGTA

  

R: CCCCGTAATGCAGAAGAAGA