Alice:LabNotes/2007-11-16: Difference between revisions
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I | Then I ran a gel on the yielded products. <br> | ||
<b>Summary of Reagents used:</b> | <b>Summary of volumes of Reagents used:</b> | ||
{| {{table}} | {| {{table}} | ||
| align="center" style="background:#f0f0f0;"|'''''' | | align="center" style="background:#f0f0f0;"|'''''' | ||
| align="center" style="background:#f0f0f0;"|''' | | align="center" style="background:#f0f0f0;"|'''well 1-2''' | ||
| align="center" style="background:#f0f0f0;"|'''well 3-4''' | |||
| align="center" style="background:#f0f0f0;"|'''well 5-8''' | |||
|- | |- | ||
| Low Mass bp Ladder ||3 ul | | Low Mass bp Ladder ||0 ul ||3 ul ||0 ul|| | ||
|- | |- | ||
| 100 bp Ladder ||3 ul | | 100 bp Ladder ||3 ul ||0 ul ||0 ul|| | ||
|- | |- | ||
| TBE buffer || 1 ul | | TBE buffer || 1 ul|| 1 ul|| 0 ul|| | ||
|- | |- | ||
| Novex 5x TBE loading dye || 1 ul | | Novex 5x TBE loading dye || 1 ul|| 1 ul|| 1 ul|| | ||
|- | |- | ||
| post-PCR sample || | | post-PCR sample || 0 ul|| 0 ul|| 3 ul|| | ||
|- | |- | ||
| | | | ||
|} | |} | ||
<b>Results:</b><br> | <b>Results:</b><br> | ||
[[Image:ZhangLab_2 2007-11-16-11hr 07min.png]] | [[Image:ZhangLab_2 2007-11-16-11hr 07min.png]] <br> | ||
Each well contain total of 5 ul of mixture of reagents. |
Revision as of 01:11, 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.
Then I ran a gel on the yielded products.
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 | 3 ul | |
Results:
File:ZhangLab 2 2007-11-16-11hr 07min.png
Each well contain total of 5 ul of mixture of reagents.