Kafatos:genomic DNA extraction from individual mosquitoes
Genomic DNA extraction from individual mosquitoes
- Put 1 mosquito into an eppendorf tube with 100ul 50% w/v Chelex 100 in water.
- Squish fly with pestle and place at 100C for 10 minutes.
- Spin at top speed in a microfuge (>14,000xg) for 1 minute.
- Transfer 10ul supernatant to new tube and add 1ul Proteinase K (0.8 units).
- Incubate at 37C for 30min.
- Incubate at 100C for 5 min.
- 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.