840:153g:Projects/project22/2012/11/29: Difference between revisions
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|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;"> Prodigiosin Production by E. Coli</span> | ||
|style="background-color: #F2F2F2" align="center"| | |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]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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== | ==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: | |||
[[Image: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. | |||
Latest revision as of 22:18, 26 September 2017
Prodigiosin Production by E. Coli | Main project page Previous entry |
Transformation
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.
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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