Alice:LabNotes/2007-11-16
PCR purification (QIAquick) on post-PCR product (reaction with newly designed primer rs1264899)
Protocol:
Add 5 volumes of Buffer PBI to 1 volume of PCR sample and mix.
Place a QIAquick spin column in a provided 2 ml collection tube.
To bind DNA, apply the sample to the QIAquick column and centrifuge for 1 min.
Discard flow-through, and place the column back into the tube.
Add 0.75 ml PE buffer to QIAquick column and centrifuge for 1 min.
Discard the flow-through and spin for additional min.
Place the column in a clean 1.5 ml microcentrifuge tube.
Add 30 ul EB buffer to center of the QIAquick membrane and centrifuge the column for 1 min to elute DNA.
Note: Store the yielded product in -20 freezer, otherwise the sample will evaporate in -4 fridge.
Then I ran a gel on the yielded products. Each well contain total of 5 ul of mixture of reagents.
Summary of volumes of Reagents used:
' | well 1-2 | well 3-4 | well 5-8 | |
Low Mass bp Ladder | 0 ul | 3 ul | 0 ul | |
100 bp Ladder | 3 ul | 0 ul | 0 ul | |
TBE buffer | 1 ul | 1 ul | 0 ul | |
Novex 5x TBE loading dye | 1 ul | 1 ul | 1 ul | |
post-PCR sample | 0 ul | 0 ul | 4 ul | |
Results: