Protocols PCRPurification

From 2008.igem.org

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