NaOH digestion of single nematodes

from Floyd, Abebe, Papert, Blaxter Molecular Ecology April 2002

Put one nematode in 20 microlitres of 0.25 M NaOH
Spin down (to make sure the nematode is fully covered by NaOH)
Optional step: freeze on dry ice or at -80 degC
Incubate overnight at 25 degC
Heat at 3 min. at 99 degC . 
Cool to room temperature. 
Spin down to collect any liquid on sides/lid of tube
Add: 	4 microlitres of 1 M HCl
	    10 microlitres 0.5 M Tris-HCl (pH 8.0)
	     5 microlitres 2% Triton X-100
Mix briefly and spin 
Heat 3 min. at 99 degC. Cool at room temperature
pH of the digest should be between 8 and 9

Store at -80 degC

Use 0.25 to 1 microlitre per 25 microlitre PCR reaction