IGEM:Virginia/2009/Notebook/VGEM2011/2011/07/27

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July 27th, 2011

  • Came into lab and realized we left ampicillin stock and miniprepped DNA at room temperature over night. Re-innoculated E. Coli from the plate. Ran an amplification PCR reaction on the three DNA segments created the day before in lab. Combined the three segments and ran a modified PCA reaction to get the final linear double stranded DNA. Analyzed results with gel electrophoresis. Found that we amplified smaller segments instead of the larger segments we wanted so when we combined the three fragments, we had very low yield.

1) ladder

2) PCR purified final construct

3) final construct

4) isolated oligos 1-20 amplified

5) isolated oligos 21-40 amplified

6) isolated oligos 41-62 amplified

7) oligos 1-20 not amplified

8) oligos 21-30 not amplified

9) oligos 41-62 not amplified



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