840:153g:Projects/project25/2012/11/29

From OpenWetWare

< 840:153g:Projects | project25 | 2012 | 11
Revision as of 17:18, 29 November 2012 by Bess E. Lippmann (Talk | contribs)
(diff) ←Older revision | Current revision (diff) | Newer revision→ (diff)
Jump to: navigation, search
Cloning of mreB from Caulobacter crescentus into E.Coli Main project page
Previous entry      

Experiment 8: Gel Electrophoresis Verification and Semester Wrap Up

Tuesday:

We digested our 12 miniprep samples with EcoR1 and Pst1. Then we ran out the samples on a gel to determine if our plasmids had the correct part inserted.

Well 1-4: samples L3D-L3A. Well 5: 100bp DNA ladder Plus. Well 6-9: samples L2D-L2A. Well 10: 100bp DNA Ladder Plus. Wells 11-14: samples L1D-L1A.

None of our plasmids seemed to have our mreB gene insert. We were expecting two bands; 1200bp and 2700bp, our insert+promoter and plasmid backbone. It looks like we got our plasmid back bone and our promoter without the gene insert (100-200bp). Well 1 had 3 bands, which suggests that it was not fully digested. The 3 bands represent super coiled, relaxed and linear DNA; while the other samples had Super coiled and linear DNA. We decided to send 2 samples (L1A and L3B) to be sequenced as a second verification method.


Thursday:

We grew 10 overnight colonies, 4 from plate L1 and 6 from plate L3, to create glycerol stocks for future use. These are 10 new colonies, so we can hopefully get our gene insert in the plasmid.


Personal tools