Maloof Lab:miniprep reagents

Buffer 1 (Cell resuspension buffer)

50mM Tris
10mM EDTA
100 ug/mL RNase A

For 100 mL
5 mL 1M Tris pH8
2 mL 0.5M EDTA
1 mL 10mg/mL RNase A
92 mL water

Buffer 2 (Cell lysis solution)

0.2M NaOH
1% SDS

For 100 mL
20 mL 1M NaOH
5 mL 20% SDS
75 mL water

Buffer 3 (Neutralization Solution)

4M guanidine hydrochloride
0.5M sodium acetate

For 100 mL
2.3g guanidine hydrochloride
4.9g potassium acetate
pH to 4.2 with glacial acetic acid
bring volume to 100 mL with water

Wash buffer

60mM potassium acetate
8.3mM Tris-HCl (pH 7.5)
40μM EDTA (pH 8)
60% ethanol [[ For 100mL]]
1.2 mL 5M potassium acetate
0.83 mL 1M Tris (pH 7.5)
8 μL 0.5M EDTA
60 mL 100% ethanol
38mL water

ADD 170 mL 95% ethanol to the wash buffer before first use!!