IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/07/29: Difference between revisions
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== | ==Tuesday 7/29== | ||
* Miniprepped the lacI and the TesA+ RBS. There was a lot of cells with both. | * Miniprepped the lacI and the TesA+ RBS. There was a lot of cells with both. | ||
* Did gel of both, and those did not turn out well. The LacI was streaked due to contamination and degradation and the the TesA + RBS was too close to the well so there was probably error in either the miniprep or the digestions of them. TesA+ RBS was digested at the x and p sites while the lacI was digested at the s and p sites. | * Did gel of both, and those did not turn out well. The LacI was streaked due to contamination and degradation and the the TesA + RBS was too close to the well so there was probably error in either the miniprep or the digestions of them. TesA+ RBS was digested at the x and p sites while the lacI was digested at the s and p sites. | ||
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| bgcolor=#cfcfcf | Reagent | | bgcolor=#cfcfcf | Reagent | ||
| bgcolor=#cfcfcf | Volume | | bgcolor=#cfcfcf | Volume | ||
| rowspan="7" | <u>Lanes+Length:</u><br> 1. 1 kb Ladder<br>3. | | rowspan="7" | <u>Lanes+Length:</u><br> 1. 1 kb Ladder<br>3. LacI = 2000<br> 5. TesA+RBS = 650<br> | ||
| rowspan="7" | [[image:7-29-14.JPG|400px]] | | rowspan="7" | [[image:7-29-14.JPG|400px]] | ||
|- | |- | ||
|| DNA(plasmid) || | || DNA (LacI plasmid) || 10.0 μL | ||
|- | |- | ||
|| 10X buffer || | || 10X buffer || 2.0 | ||
|- | |- | ||
|| | || PstI || 1.0 | ||
|- | |- | ||
|| | || SpeI || 1.0 | ||
|- | |- | ||
|| dH<sub>2</sub>O || | || dH<sub>2</sub>O || 6.0 | ||
|- | |- | ||
|| || | || || 20.0 μL --> 37°C/ ~1 Hr. | ||
|} | |} | ||
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Revision as of 14:41, 29 July 2014
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Tuesday 7/29
Had to restart those. Did the seeds of both, and re-streaked the TesA +RBS from plate 4, colony 3.
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