Component	Amount (ml)	Final concentration
Tris, pH 8.0, 1M	1.2	4.8 mM
Tris, pH 7.5, 1M	1.2	4.8 mM
MgCl2, 1M	0.125	0.5 mM
CaCl2, 1M	0.110	0.44 mM
Dtt, 0.1M	0.025	10 μM
Proteinase K, 300 U ml−1	2.5	3 U ml−1
Triton X-114, 10% wt/vol	2.5	0.1% (wt/vol)
UltraPure Water	242
Total	250