Team:CSU Fort Collins/Notebook/Breakdown/Aug
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<h4>Tuesday, August 5</h4> | <h4>Tuesday, August 5</h4> | ||
- | Reran PCR with correct primer using previously isolated E. coli genome. | + | Reran PCR with correct primer using previously isolated <u>E. coli</u> genome. |
<h4>Wednesday, August 6</h4> | <h4>Wednesday, August 6</h4> | ||
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<h4>Friday, August 8</h4> | <h4>Friday, August 8</h4> | ||
- | Discussed with Sun why the PCR was not working. Discovered that when finding the melting temperatures we had not used the specifications of the polymerase we were using. Reran the PCR using a gradient around the new melting temperature. We used both colonies and previously isolated E. coli genome in different replicates for our template DNA. Ran a gel to determine success. The conditions which gave us the required 1.7 kb strand were 61 ºC and 1.7 minute extension step for the first five cycles and 72 ºC and 1.7 minute extension step for the last 30 cycles. The template DNA that worked came from the previously isolated E. coli genome. | + | Discussed with Sun why the PCR was not working. Discovered that when finding the melting temperatures we had not used the specifications of the polymerase we were using. Reran the PCR using a gradient around the new melting temperature. We used both colonies and previously isolated <u>E. coli</u> genome in different replicates for our template DNA. Ran a gel to determine success. The conditions which gave us the required 1.7 kb strand were 61 ºC and 1.7 minute extension step for the first five cycles and 72 ºC and 1.7 minute extension step for the last 30 cycles. The template DNA that worked came from the previously isolated <u>E. coli</u> genome. |
<h4>Monday, August 18</h4> | <h4>Monday, August 18</h4> | ||
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<h4>Tuesday, August 19</h4> | <h4>Tuesday, August 19</h4> | ||
- | Transformed into E. coli to amplify the plasmid. | + | Transformed into <u>E. coli</u> to amplify the plasmid. |
<h4>Wednesday, August 20</h4> | <h4>Wednesday, August 20</h4> |
Revision as of 03:09, 14 October 2014
Breakdown Daily Notes
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July
Lac Promoter and FadD Assembly
August
PLR, PFL, and PFL2 Assembly
September
Scientists vs. the PCR Machine