Haynes Lab:Notebook/Jan/2015/03/20: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Line 20: | Line 20: | ||
# BD006 a | # BD006 a | ||
# BD006 b | # BD006 b | ||
* Digests (Fermentas FD) | |||
{| {{table}} cellspacing="3" <!-- Digest rxn. table --> | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <!-- [[Image:GelImage.jpg|270px|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] --> | |||
|- | |||
| DNA (plasmid) || up to 25 μL | |||
|- | |||
| 10x buffer || 3.0 | |||
|- | |||
| enzyme 1 || 1.0 | |||
|- | |||
| enzyme 2 || 1.0 | |||
|- | |||
| dH<sub>2</sub>O || --- | |||
|- | |||
| || 30 μL --> 37°C/ ~30 min. | |||
|} | |||
* Conclusions | |||
** MV2 is correct | |||
** BD004 - most likely missing the D11.12 fragment. Sequence results from Meli'sa's past work agrees with the short fragment size on the gel. | |||
** BD006 is correct | |||
'''Assemblies''' | |||
# BD006/MV2: BD006(E/S)/4000 + MV2(E/X)/4500 | |||
* Gel purification: Sigma Gel extraction kit, elute and back-elute with 40 μL elution buffer | |||
** MV2 (E/X) | |||
** BD006 (E/S) | |||
* Measure conc.'s | |||
{| {{table}} cellspacing="3" <!-- [DNA] table --> | |||
|- bgcolor=#cfcfcf | |||
| Sample || OD260 || 260/280 || ng/μL | |||
|- | |||
| 1. Digested part (a/b) || --- || --- || --- | |||
|- | |||
| 2. Digested part (c/d) || --- || --- || --- | |||
|} | |||
Revision as of 13:12, 21 March 2015
Project name | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | |||||||||||||||||||||||||||
03/20/15
Plasmids - minipreps & digests
|