Alice:LabNotes/2007-11-16

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Revision as of 01:13, 1 December 2007 by >Zsakura2
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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:

File:ZhangLab 2 2007-11-16-11hr 07min.png