User:Tara K. Luckau/Notebook/Team ConGen/2010/11/08

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 * style="background-color: #BCED91"|[[Image:owwnotebook_icon.png|128px]] Tara's Lab Notebook
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 * style="background-color: #9DB68C" align="center"|  |Main project page


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Cast Gel

 * 270µL 1x TAE + 5.4g agarose
 * hot plate + stir bar
 * 27µL Gel Red (added when agarose turned clear but still hot, used stir bar)

Load Gel

 * 5µL NanoPure H2O + 5µL sample
 * (to conserve loading dye)
 * (don't need loading dye in every sample since it's in ladder)
 * 10µL ladder / loading dye mix
 * [[Image:20101108 GelSchematic.jpg|800 px]]

Run Gel

 * 130C, 2 hrs

Image Gel

 * UGH!
 * [[Image:20101108_Top.TIF|300 px]]   [[Image:20101108_TopZoom.TIF|300 px]]
 * [[Image:20101108_Bottom.TIF|300 px]]   [[Image:20101108_BottomZoom.TIF|300 px]]
 * 3 faint bands

Results

 * Very faint bands at E8, E10, E12, corresponding to DNA concentration of 1x, Buffer G, Temperatures 61, 63, 65
 * around 150 bp band

Next Steps

 * there's obviously a problem:
 * reagent sucks (made wrong, not stored properly, expired)
 * inhibition
 * Start weeding out the problem
 * check math on dNTPs, ladder (too bright?)
 * try different Taq?
 * try Scun22 primers


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