Haynes Lab:Notebook/Engineering PC-TFs/2014/04/18: Difference between revisions
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(Autocreate 2014/04/18 Entry for Haynes_Lab:Notebook/Engineering_PC-TFs) |
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==Summary== | ==Summary== | ||
* | 4/16 | ||
*Cloned mv9cmv construct into DH5a-T cells using standard procedure. Incubated overnight. | |||
4/17 | |||
* Removed plates from incubator. | |||
{| {{table}} border="1" cellspacing="3" <!-- Plates. table --> | |||
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| rowspan="7" | [[Image:2014-04-17 15.36.21.jpg|400px|Hover name]]; | |||
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| rowspan="7" | [[Image:2014-04-17 15.36.39.jpg|400px|Hover name]]<br>; | |||
|- valign="top" | |||
|} | |||
*Plate 1 is the experimental plate with growth. Plate 2 is the negative control and had no growth (debris is visible underneath the plate). | |||
4/18 | |||
*Grew up liquid cultures from prior night. | |||
*Miniprepped cultures. | |||
{| {{table}} | |||
|- bgcolor=#cfcfcf | |||
| Plasmid || OD260 || OD260/280 || ng/μL | |||
|- | |||
| 1. Colony 1 || 0.202 || 1.905 || 162 | |||
|- | |||
| 2. Colony 2 || 0.253 || 1.92 || 203 | |||
|} | |||
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{| {{table}} border="1" cellspacing="3" | |||
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|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. CMV = 588bp <br> | |||
|- | |||
| DNA(CMV/MV9) || 4.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| NotI || 1.0 μL | |||
|- | |||
| NruI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 7.5 μL | |||
|- | |||
| || 15 μL --> 37°C/ 15 min. | |||
|} | |||
*Verified for the sake of not wasting money on sequencing. | |||
{| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table --> | |||
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| rowspan="7" | [[Image:2014-04-18 13.13.10.jpg|400px|Hover name]]<br>15 μL/lane; 1% agarose; cut the brightest band furthest down at ~ 588bp; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | |||
|} | |||
*Expected base pair length: 643 | |||
__NOTOC__ | __NOTOC__ |
Revision as of 14:42, 18 April 2014
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Summary4/16
4/17
4/18
|
Reagent | Volume | Expected: 1. CMV = 588bp |
DNA(CMV/MV9) | 4.0 μL | |
10X buffer | 1.5 μL | |
NotI | 1.0 μL | |
NruI | 1.0 μL | |
dH2O | 7.5 μL | |
15 μL --> 37°C/ 15 min. |
- Verified for the sake of not wasting money on sequencing.
15 μL/lane; 1% agarose; cut the brightest band furthest down at ~ 588bp; Ladder |
- Expected base pair length: 643