Alice:LabNotes/2007-11-16: Difference between revisions
>Zsakura2 (New page: PCR purification (QIAquick) on post-PCR product (reaction with newly designed primer rs1264899)<br> <b>Protocol:</b> <br> Add 5 volumes of Buffer PBI to 1 volume of PCR sample and mix.<br>...) |
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[[Image:ZhangLab_2 2007-11-16-11hr 07min.png]] | [[Image:ZhangLab_2 2007-11-16-11hr 07min.png]] |
Revision as of 00:48, 1 December 2007
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.
I also ran another gel on the 6 samples that failed on DNA sequencing.
Summary of Reagents used:
' | volume |
Low Mass bp Ladder | 3 ul |
100 bp Ladder | 3 ul |
TBE buffer | 1 ul |
Novex 5x TBE loading dye | 1 ul |
post-PCR sample | 4 ul |