Protocols ColonyPCR

Procedure 1  Add 5 volumes (~250mL) of Buffer PBI to 1 volume (~50mL) of the PCR sample mix 2  Check that the color of the mixture is yellow (similiar to Buffer PBI w/o PCR sample) If the color is orange or violet, add 10ul of 3M sodium acetate, pH 5.0 and mix 3  Place a QIAquick spin column in a 2mL collection tube (Purple Column) 4  To bind DNA, apply the sample to the QIAquick column and centrifuge for 30-60s 5  Discard flow-through. Place the QIAquick column back into the same tube 6  To wash, add 0.75mL Buffer PE to the QIAquick column and centrifuge for 30-60s 7  Discard flow-through and place the QIAquick column back in the same tube. Centrifuge the column for an additional 1 minute to evaporate ethanol 8  Place QIAquick column in a clean 1.5mL microcentrifuge tube 9  To elute DNA, add 50ul Buffer EB (or 2mMTris) or water to the center of the QIAquick membrane and centrifuge the column for 1 minute. Alternatively, for increased DNA concentration, add 30ul elution buffer to the center of the QIAquick membrane, let the column stand for 1 minute and then centrifuge. Maximum elution efficiency with pH 7.0-8.5 10  Store DNA at -20*C

Protocols