Difference between revisions of "840:153g:Projects/project22/2012/11/29"

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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;"> Prodigiosin Production by 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>}}
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|style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
 
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==Entry title==
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==Transformation==
* Insert content here...
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* Before break, we began the process of transformation using the pGEM-T Easy Vector kit. On Tuesday we resumed transformation and transformed our PCR product in DH5alpha cells and grew them on ampicillin plates.  
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* Plate Results:
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  3Amp: 2 colonies (only one was present at the time of picking)
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  2Amp: 0 colonies
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  1Amp: 0 colonies
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  +Amp: 20 colonies
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  -Amp: 0 colonies
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  -NoAmp: plate completely covered.
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* Since out of all of our sample plates, plate 3 was the only to produce a single colony, it was picked and incubated in 5 mL of luria broth and 5 ul of ampicillin at 37 degrees Celsius and 220 rpm. Growth was observed in all of the test tubes(1-5 positive controls and Ampicillin plate 3 colony).
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* Next, we used the GeneJet Plasmid Miniprep Kit (Lot: 00038294) to purify the plasmid, and ran gel electrophoresis on the product. Our results are as follows:
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[[Image:29.11.12_small.png‎]]
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The top band on the ladder (first lane) represents 3000 b.p. As you can see, we have a bright band above this line (lane 2) which corresponds to the size of our T-vector (~3200 b.p.). We do not, however, see a bright line at ~1500 b.p. which we would expect with successful transformation of our gene (pigI is 1473 b.p.). To confirm these results, we will send off our DNA product for sequencing.
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Latest revision as of 21:18, 26 September 2017

Owwnotebook icon.png Prodigiosin Production by E. Coli Report.pngMain project page
Resultset previous.pngPrevious entry      

Transformation

  • Before break, we began the process of transformation using the pGEM-T Easy Vector kit. On Tuesday we resumed transformation and transformed our PCR product in DH5alpha cells and grew them on ampicillin plates.
  • Plate Results:
  3Amp: 2 colonies (only one was present at the time of picking)
  2Amp: 0 colonies
  1Amp: 0 colonies
  +Amp: 20 colonies
  -Amp: 0 colonies
  -NoAmp: plate completely covered.
  • Since out of all of our sample plates, plate 3 was the only to produce a single colony, it was picked and incubated in 5 mL of luria broth and 5 ul of ampicillin at 37 degrees Celsius and 220 rpm. Growth was observed in all of the test tubes(1-5 positive controls and Ampicillin plate 3 colony).
  • Next, we used the GeneJet Plasmid Miniprep Kit (Lot: 00038294) to purify the plasmid, and ran gel electrophoresis on the product. Our results are as follows:

29.11.12 small.png

The top band on the ladder (first lane) represents 3000 b.p. As you can see, we have a bright band above this line (lane 2) which corresponds to the size of our T-vector (~3200 b.p.). We do not, however, see a bright line at ~1500 b.p. which we would expect with successful transformation of our gene (pigI is 1473 b.p.). To confirm these results, we will send off our DNA product for sequencing.