Alice:LabNotes/2007-11-16: Difference between revisions

From ZhangLabWiki
Jump to navigation Jump to search
>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>...)
 
>Zsakura2
No edit summary
Line 30: Line 30:
|
|
|}  
|}  
<b>Results:</b>
<b>Results:</b><br>
[[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

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