Editing Plasmid buffers (section)

== <u>'''Maxiprep'''</u> ==

<br/>'''QC buffer''' (wash buffer)

1.0M NaCl<br/>50 mM MOPS pH 7.0<br/>15% isopropanol

to make 2 L<br/>666 ml 3M NaCl<br/>200 ml 0.5 M MOPS pH 7.0<br/>300 ml EtOH<br/>dH2O to 2 liters<br/>check the pH and adjust to 7.0 (it should be close) with NaOH or HCl

'''QF buffer''' (elution buffer)

1.25M NaCl<br/>50 mM Tris-HCl pH 8.5<br/>15% isopropanol

to make 2 L<br/>833 ml 3M NaCl<br/>100 ml 1M Tris pH 8.5<br/>300 ml EtOH<br/>dH2O to 2 liters<br/>check the pH and adjust to 8.5 (it should be close) with NaOH or HCl<br/>store QC and QF in tightly capped containers at room temp.



'''Buffer QN'''

1.6M NaCl<br/>50 mM MOPS pH 7.0<br/>15% isopropanol



'''Buffer FWB2'''

1M potassium acetate, pH 5.0



'''QBT buffer'''

750 mM NaCl<br/>50 mM MOPS pH 7.0<br/>15% isopropanol<br/>0.15% Triton X-100

to make 2 L:<br/>500 ml 3M NaCl<br/>200 ml 0.5 M MOPS pH 7.0<br/>300 ml EtOH<br/>3 ml Triton X-100<br/>dH2O to 2 liters

Buffer QBT (equilibration buffer)



'''P1 buffer'''<br/>1M Tris HCl pH 8.0 2.5 ml<br/>0.5 M EDTA pH 8.0 1 ml<br/>10 mg/ml RNase A, boiled 0.5 ml<br/>dH2O 46 ml<br/>50ml total<br/><br/>'''P2 buffer'''<br/>2M NaOH 5 ml<br/>20% SDS 2.5 ml<br/>dH2O 42.5 ml<br/>50 ml total<br/>Apparently this solution can go bad due to oxidation: keep the container tightly capped.<br/><br/>'''P3 buffer'''<br/>potassium acetate 25 g<br/>glacial acetic acid 13 ml<br/>H2O 87 ml<br/>100 ml total<br/>final pH should be ~5.5