Fecal DNA Extraction Protocol
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Corresponding Organization :
Other organizations : Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes, Institut de Recherche pour le Développement, Agropolis International, Méditerranée Infection Foundation
Protocol cited in 1 other protocol
Variable analysis
- Removal of the outer layer of fecal bulk (approximately 1–2 mm)
- Use of the inner part of the fecal bulk for extraction
- DNA extracted from the fecal samples
- Use of sterile scalpel and polystyrene tweezers
- Avoidance of possible contamination with soil organisms and/or the risk of egg deposition by some flies
- Use of a modified version of the Qiagen fecal procedure (QIAamp DNA Tissue Kit, Qiagen Inc., Germany)
- Use of a 200-mg aliquot of each fecal sample
- Use of a 2-ml tube containing 200 mg of a mixture of 0.1-, 0.5-, and 2-mm zirconium beads and 1.5 ml of ASL buffer (Qiagen)
- Bead-beating at 3200 rpm for 90 seconds
- Heating at 95°C for 10 minutes
- Suspension of the final pellet in 180 μl of tissue lysis buffer and incubation with proteinase K for 2 hours at 55°C
- Following the manufacturer's recommendations for the purification and elution of DNA
- Not explicitly mentioned
- Not explicitly mentioned
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