Difference between revisions of "840:153g:Projects/project25/2012/11/29"

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project Name</span>
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Cloning of mreB from ''Caulobacter crescentus'' into ''E.Coli''</span>
 
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
 
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
 
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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.
  
  

Revision as of 14:18, 29 November 2012

Owwnotebook icon.png Cloning of mreB from Caulobacter crescentus into E.Coli <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>

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.

2012-11-29 15.04.08.jpg

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.