840:153g:Projects/project25/2012/11/29: Difference between revisions
(Autocreate 2012/11/29 Entry for 840:153g:Projects/project25) |
No edit summary |
||
| (5 intermediate revisions by the same user not shown) | |||
| Line 1: | Line 1: | ||
{|{{table}} width="800" | {|{{table}} width="800" | ||
|- | |- | ||
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> | |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> </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> </html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}} | ||
|- | |- | ||
| colspan="2"| | | colspan="2"| | ||
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | ||
== | ==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. | |||
[[Image:2012-11-29 15.04.08.jpg|400px]] | |||
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. | |||
Revision as of 14:18, 29 November 2012
 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> </html> |
Experiment 8: Gel Electrophoresis Verification and Semester Wrap UpTuesday: 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.
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.
| |
