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

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I also ran another gel on the 6 samples that failed on DNA sequencing.<br>
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;"|'''volume'''
| 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 || 4 ul
| 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.