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 Tara's Lab Notebook
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Gel Scun2 F-R with DNA Gradient
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
Run Gel
Image Gel
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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Tara's Lab Notebook
