Kafatos:genomic DNA extraction from individual mosquitoes

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Genomic DNA extraction from individual mosquitoes

  1. Put 1 mosquito into an eppendorf tube with 100ul 50% w/v Chelex 100 in water.
  2. Squish fly with pestle and place at 100C for 10 minutes.
  3. Spin at top speed in a microfuge (>14,000xg) for 1 minute.
  4. Transfer 10ul supernatant to new tube and add 1ul Proteinase K (0.8 units).
  5. Incubate at 37C for 30min.
  6. Incubate at 100C for 5 min.
  7. Use 0.5ul as template in a 10ul PCR reaction.

Reagents (from Sigma):
Chelex 100 (C7901-25G) sodium form chelating resin (iminodiacetic acid)

Proteinase K (P4850) from Tritirachium album, for molecular biology,
>800 units/mL, buffered aqueous glycerol solution, DNAse-, Nickase- and RNAse- free.