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

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==Title here==
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==Experiment 8: Gel Electrophoresis Verification and Semester Wrap Up==
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'''Tuesday''':
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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.
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[[Image:2012-11-29 15.04.08.jpg|400px]]
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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.
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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.
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'''Thursday''':
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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.

Current revision

Cloning of mreB from Caulobacter crescentus into E.Coli Main project page
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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.


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