Minipreps (Quiagen, Inc) are used to purify DNA out of bacterial cultures. Variants of minipreps can be used to clean DNA after a PCR or extract it from an agarose gel matrix
P1 buffer
1M Tris HCl pH 8.0 2.5 ml
0.5 M EDTA pH 8.0 1 ml
10 mg/ml RNAseA, boiled 0.5 ml
dH2O 46 ml
50ml total
P2 buffer
2M NaOH 5 ml
20% SDS 2.5 ml
dH2O 42.5 ml
50 ml total
Apparently this solution can go bad due to oxidation: keep the container tightly capped.
P3 buffer
potassium acetate 25 g
glacial acetic acid 13 ml
H2O 87 ml
100 ml total
final pH should be ~5.5
QC buffer
666 ml 3M NaCl200 ml 0.5 M MOPS pH 7.0
300 ml EtOH
dH2O to 2 liters
check the pH and adjust to 7.0 (it should be close) with NaOH or HCl
QF buffer
833 ml 3M NaCl
100 ml 1M Tris pH 8.5
300 ml EtOH
dH2O to 2 liters
check the pH and adjust to 8.5 (it should be close) with NaOH or HCl
store QC and QF in tightly capped containers at room temp.
QBT buffer
500 ml 3M NaCl
200 ml 0.5 M MOPS pH 7.0
300 ml EtOH
3 ml Triton X-100
dH2O to 2 liters
To make these, you will need the following stock solutions:
0.5 M MOPS pH 7.0
209.27 g MOPS (use the free acid form that USB sells)
~750 ml dH20
pH to 7.0 with 10 N NaOH
make up to 2 liters with dH20
3M NaCl
350.6 g Nacl
make up to 2 liters with dH20
1 M Tris pH8.5
121.1 g Tris base
~750 ml dH20
pH to 8.5 with concentrated HCl (takes ~20 ml)
make up to 1 liter with dH20
Note: You can store these solutions at room temp.