Worm Lysis

Worm lysis can be performed on a single worm or a pool of worms. For a pool, 50-60 worms should be enough.

Worm Lysis Solution (10ml):

500ul 1M KCl

100ul 1M Tris pH8.3

25ul 1M MgCl₂

45ul NP40

45ul Tween 20

50ul 2% gelatin

ddH₂O to 10ml

Each time a worm lysis is done, take a 950ul aliquot of the worm lysis solution and add 50ul Proteinase K (stock solution at 20mg/ml).

In a 0.5ml tube, use 50-60 ul for a pool of 50-60 worms. Use up to 30 or 40ul for a single worm.

Freeze the tube at -80°C. This is a freeze-crack step, which will help liberate the DNA.

Incubate the tube at 60°C for 60 minutes, followed by a 95°C incubation for 15 minutes. Cool to 4°C by placing on ice. Alternatively, this whole cycle is on the MJ PTC-100 thermocycler in a program called "60-95-4."

The lysed worm solution is now ready to use for PCR, or other applications.

For a larger, cleaner genomic DNA stock, see the "Genomic DNA Prep" protocol.