Molecular Characterization of Mosquito Species

Genomic DNA was extracted from F₁ individual mosquitoes (n = 29) representing all adult morphological variations, using a phenol–chloroform extraction method [18 (link)]. The ITS2 region [19 (link)], D3 region [20 (link)], white gene [21 (link)], mitochondrial COI [22 (link)], and Cyt-b [23 (link)] were amplified in a 25 µL reaction mixture. The reaction and cyclic conditions for each PCR are shown in Table 1. Amplified PCR products were purified using QIAquick PCR product purification kit (Qiagen, Hilden, Germany) and sequenced bi-directionally at Macrogen Inc., Seoul, South Korea.

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Jayatunga D.P., Harischandra I.N., Chandrasekharan N.V, & de Silva N.K. (2018). Alterations and Interchange of Morphometric Characters in Different Life Cycle Stages with Reference to Genomic Variations of Anopheles subpictus (Diptera; Culicidae) Sibling Species Complex in Sri Lanka. Insects, 9(3), 89.

Publication 2018

QIAquick PCR product purification kit

Adult Chloroform Gene Genomic Mitochondrial Mosquitoes Phenol

Corresponding Organization : University of Sri Jayewardenepura

Other organizations : University of Colombo

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Variable analysis

independent variables

Genomic DNA extraction method (phenol–chloroform extraction)

dependent variables

Nucleotide sequence of ITS2 region
Nucleotide sequence of D3 region
Nucleotide sequence of white gene
Nucleotide sequence of mitochondrial COI
Nucleotide sequence of Cyt-b

control variables

Adult mosquito morphological variations (all variations represented)
Sample size (n = 29 F1 individual mosquitoes)
PCR reaction mixture (25 µL)
PCR reaction and cyclic conditions (as shown in Table 1)
Purification of amplified PCR products (using QIAquick PCR product purification kit)
Sequencing method (bi-directional sequencing at Macrogen Inc., Seoul, South Korea)

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