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% SDA

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) 40um EDTA (pH 8) 60% ethanol

For 100mL 1.2 mL 5M potassium acetate 0.83 mL 1M Tris (pH 7.5) 8 ul 0.5M EDTA 60 mL 100% ethanol 38mL water

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