Alice:LabNotes/2007-11-26: Difference between revisions

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Run PCR purification with post-PCR product from the 6 failed sample from DNA sequencing, which was the original sample from well #1,2,3,12,14,16. The method used this time is different from the method using column. This method is more old school way that takes longer but allows greater %yield. It is good if one wants to preserve greater amount of DNA.<br>
Purify the post-PCR samples with EXOsap-IT, and then run a gel to check if all the reaction worked fine.
Each well contain total of 5 ul of mixture of reagents.<br>
Each well contain total of 5 ul of mixture of reagents.<br>
<b>Summary of volumes of Reagents used:</b>
<b>Summary of volumes of Reagents used:</b>

Revision as of 09:42, 2 December 2007

Run PCR purification with post-PCR product from the 6 failed sample from DNA sequencing, which was the original sample from well #1,2,3,12,14,16. The method used this time is different from the method using column. This method is more old school way that takes longer but allows greater %yield. It is good if one wants to preserve greater amount of DNA.
Purify the post-PCR samples with EXOsap-IT, and then run a gel to check if all the reaction worked fine. Each well contain total of 5 ul of mixture of reagents.
Summary of volumes of Reagents used:

' well 1 well 2 well 3 well 4 well 5 well 6 well 7 well 8
Low Mass bp Ladder 3 ul 0 ul 0 ul 0 ul 0 ul 0 ul 0 ul 0 ul
100 bp Ladder 0 ul 3 ul 0 ul 0 ul 0 ul 0 ul 0 ul 0 ul
TBE buffer 1 ul 1 ul 0 ul 0 ul 0 ul 0 ul 0 ul 0 ul
Novex 5x TBE loading dye 1 ul 1 ul 1 ul 1 ul 1 ul 1 ul 1 ul 1 ul
post-PCR sample 0 ul 0 ul 4 ul 4 ul 4 ul 4 ul 4 ul 4 ul

Results:
File:ZhangLab 2 2007-11-26 13hr 17min-2-.png
This could have been a rather good gel if the focus of the gel imaging machine could be adjusted to be either further away of closer to the level of samples. The gel was stained with SYBGold for 15 min. Even though the gel is fuzzy, but it is easy to tell that the intensity seems to be desirable, since all the bands are visible. Well #3, #4, and #6 shows weaker intensity maybe because the reaction didn't shoot up until very late stage, so they are not as desirable as the other three. File:ZhangLab 2 2007-11-26 14hr 29min.png
File:ZhangLab 2 2007-11-26 14hr 36min.png