protocols

DNA preparation for PCR product less than 500 bp

Lysis buffer:

	Final concentration	Stock concentration	For 10 ml mix
Tris pH8.8	50 mM	1 M	500 ul
EDTA pH 8.0	1 mM	0.5 M	20 ul
Tween	0.5%	10%	500 ul
H2O			9000 ul

Add Proteinase K before use.
For tail piece 0.2mm long or ear piece, add 50ul buffer plus 1.5 ul of 20 mg/ml (30ug) Proteinase K stock solution
10 samples: 550 ul lysis buffer + 16.5 ul PK 20 mg/ml
you can use 0.5 ml PCR tubes for digestion and heat in a PCR machine.
Lysis-
55C, O/N , then heat inactive at 100C for 10 minutes. Cool down on bench top.
For 20ul PCR reaction, use 0.8ul tail DNA.