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Protocols PCRPurification
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RAUL CUERO (Talk | contribs) (New page: '''Procedure'''<br> '''1''' Add 5 volumes (~250mL) of Buffer PBI to 1 volume (~50mL) of the PCR sample mix <br> '''2''' Check that the color of the mixture is yellow (similiar to B...) |
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Latest revision as of 17:28, 29 October 2008
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
