Haynes Lab:Notebook/Jan/2015/04/10: Difference between revisions
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(Autocreate 2015/04/10 Entry for Haynes_Lab:Notebook/Jan) |
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== | ==04/10/15== | ||
* | * Minipreps - BD004/MV2 (6x 5mL cultures) | ||
---- | |||
* Sigma GenElute kit. | |||
** Elute & back-elute w/ 70 μL elution sln. | |||
# BD004/MV2 a | |||
# BD004/MV2 b | |||
# BD004/MV2 c | |||
# BD004/MV2 d | |||
# BD004/MV2 e | |||
# BD004/MV2 f | |||
* Restriction Digest check (Fermentas FD) | |||
** Cut with ApaLI (E/S insertion got rid of BB RFC23 cut sites) | |||
{| {{table}} cellspacing="3" <!-- Digest rxn. table --> | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | Expected:<br>* BD004/MV2 = 7108, 1243, 288<br>* MV2 only = 2998, 1243, 288<br>* BD004/V0120 = 6905, 497 | |||
| rowspan="7" | [[Image:KAH032815_gel1.jpg|270px|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | |||
|- | |||
| DNA (plasmid) || 3.0 μL | |||
|- | |||
| 10x buffer || 1.5 | |||
|- | |||
| ApaLI || 1.0 | |||
|- | |||
| dH<sub>2</sub>O || 9.5 | |||
|- | |||
| || 15 μL --> 37°C/ ~10 min. | |||
|} | |||
* Conclusions | |||
** Success! All four look like BD006/MV2 | |||
** Submit for sequencing next week | |||
* Miniprep conc.'s | |||
{| {{table}} cellspacing="3" <!-- [DNA] table --> | |||
|- bgcolor=#cfcfcf | |||
| Sample || OD260 || 260/280 || ng/μL | |||
|- | |||
| 1. BD006/MV2 1-a || 0.274 || 1.44 || 274.10 | |||
|- | |||
| 2. BD006/MV2 1-b || 0.299 || 1.93 || 298.68 | |||
|- | |||
| 3. BD006/MV2 2-a || 0.331 || 1.93 || 330.69 | |||
|- | |||
| 4. BD006/MV2 2-b || 0.306 || 1.90 || 305.94 | |||
|} | |||
Revision as of 14:54, 10 April 2015
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04/10/15
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