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Alice:LabNotes/2007-11-28
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Since the gel from yesterday didn't work out, so some more sample (from well #1,2,3,12,14,16) were purified with EXOsap-IT. Then I ran a agrose gel. Each well contain total of 5 ul of mixture of reagents.<br> <b>Summary of volumes of Reagents used:</b> {| {{table}} | align="center" style="background:#f0f0f0;"|'''''' | align="center" style="background:#f0f0f0;"|'''well 1''' | align="center" style="background:#f0f0f0;"|'''well 2''' | align="center" style="background:#f0f0f0;"|'''well 3''' | align="center" style="background:#f0f0f0;"|'''well 4''' | align="center" style="background:#f0f0f0;"|'''well 5''' | align="center" style="background:#f0f0f0;"|'''well 6''' | align="center" style="background:#f0f0f0;"|'''well 7''' | align="center" style="background:#f0f0f0;"|'''well 8''' |- | Low Mass bp Ladder ||3 ul ||0 ul ||0 ul||0 ul||0 ul||0 ul||0 ul||3 ul|| |- | TBE buffer || 1 ul|| 1 ul|| 1 ul|| 1 ul|| 1 ul|| 1 ul|| 1 ul|| 1 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 ||3 ul ||3 ul ||3 ul ||3 ul ||3 ul ||3 ul ||0 ul || |- | |} <b>Results:</b><br> [[Image:ZhangLab_2 2007-11-28 16hr 08min.png]]<br> The gel finally worked! As we can tell from the image, there was no fuzzy edge or blurry effect, so the focus of the machine is desirable. However, the gel is not bright enough for quantization. This is probably because the gel didn't stain for long enough. I used 40uL SYBEsafe in 400mL TBE buffer, which is from the Invitrogen website instruction.
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