840:153g:Projects/project22/2012/11/15
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Current revision (19:14, 15 November 2012) (view source) |
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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 in 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>}} | ||
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| - | == | + | ==PCR DNA Gel Extraction and Purification and the Beginning of T Vector Ligation== |
| - | + | On Tuesday we ran a gel electrophoresis check on our PCR and it yielded these results: | |
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| + | [[Image:13.11.12_small.png]] <--- Note the bands at the approximate 1500 b.p. (above the second bright line on the markers in lanes 9 and 18). Also note the functional positive control in lane 10 | ||
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| + | Based on these results, we believe we have amplified our target gene (pigI) so we extracted the DNA from the gel and purified it with a GeneJet Gel Extraction kit (Lot 00040498). We ran a gel electrophoresis check on the sample obtained from gel extraction and it yielded these results: | ||
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| + | [[Image:15.11.12_small.png]] <-- Note the DNA at approximately 1500 b.p. (slightly above the second very bright line, which represents 1000 b.p. - slightly blurred in this picture - on the marker in lane 6) | ||
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| + | This gel confirms our belief that we have our target gene, so we proceeded to the next step which is transformation and cloning of our DNA using a T-vector. To this end, we started the T Vector Ligation, using the Promega pGEM-T Easy Vector Ligation "Cloning PCR" protocol and will begin transformation on next Tuesday. | ||
Current revision
 Prodigiosin Production in E. Coli
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PCR DNA Gel Extraction and Purification and the Beginning of T Vector LigationOn Tuesday we ran a gel electrophoresis check on our PCR and it yielded these results:
Based on these results, we believe we have amplified our target gene (pigI) so we extracted the DNA from the gel and purified it with a GeneJet Gel Extraction kit (Lot 00040498). We ran a gel electrophoresis check on the sample obtained from gel extraction and it yielded these results:
This gel confirms our belief that we have our target gene, so we proceeded to the next step which is transformation and cloning of our DNA using a T-vector. To this end, we started the T Vector Ligation, using the Promega pGEM-T Easy Vector Ligation "Cloning PCR" protocol and will begin transformation on next Tuesday.
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