Home › Analytical Chem › A Simple Chelex Protocol for DNA Extraction from Anopheles spp.
Analytical Chem JoVE (Open Access) Citable · DOI

A Simple Chelex Protocol for DNA Extraction from Anopheles spp.

DOI: 10.3791/3281-v
What you'll learn
  • ✓ Extract quality DNA from Anopheles mosquito specimens using Chelex resin
  • ✓ Perform molecular identification of Anopheles sibling species by PCR
  • ✓ Genotype Plasmodium falciparum parasites from mosquito tissues
Protocol

A rapid and affordable way to extract quality malaria parasite and vector DNA from mosquito specimens is described. Capitalizing on chelating properties of Chelex resin, the simple method enables genotyping of malaria parasites in mosquito mid-gut and salivary gland phases, as well as molecular identification of the Anopheles sibling species by PCR.

Difficulty
beginner
Total time
~2-3 hours per 10 specimens (including dissection, extraction, and PCR)
Model organism
Anopheles spp. (malaria vector mosquitoes)
Biosafety
BSL-2

Steps

1
Dissect mosquito specimens and isolate tissues

Prepare mosquito specimens by dissecting mid-gut and salivary gland tissues. These tissues will be used for subsequent DNA extraction and parasite genotyping.

▶ 01:33
2
Extract DNA using Chelex resin protocol

Incubate dissected mosquito tissues with Chelex resin to chelate inhibitory ions and extract quality DNA. This simple method produces DNA suitable for PCR amplification without additional purification steps.

▶ 02:28
3
Perform PCR genotyping and species identification

Conduct PCR assays using extracted DNA to genotype Plasmodium falciparum parasites and perform molecular identification of Anopheles species using species-specific primers.

▶ 04:03
4
Analyze and interpret PCR assay results

Evaluate PCR products via gel electrophoresis to confirm successful parasite genotyping and species identification based on expected amplicon sizes.

▶ 05:25
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