User:Karmella Haynes/Notebook/BioBrick cloning/2013/04/01: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
Line 27: Line 27:
| bgcolor=#cfcfcf | Volume  
| bgcolor=#cfcfcf | Volume  
| rowspan="7" | <u>Expected:</u><br>1. BB 1 = size<br>2. BB2 = size<br>
| rowspan="7" | <u>Expected:</u><br>1. BB 1 = size<br>2. BB2 = size<br>
| rowspan="7" | [[Image:KAH040113.jpg|200px|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
| rowspan="7" | [[Image:KAH040113_gel1.jpg|200px|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
|-
|-
| DNA(plasmid) || 3.0 μL
| DNA(plasmid) || 3.0 μL

Revision as of 06:24, 2 April 2013

Karmella's BioBrick Cloning <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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

04/01/13

  • Check MV9 clones


MV9 clones

  • Picked colonies from last cloning attempt. Streaked on LB amp agar.
  • Grew up 2 mL cultures overnight and did minipreps.


Minipreps

  • Check with S/X digests
    • S present in upstream cloning sites, X inside the new insert
Reagent Volume Expected:
1. BB 1 = size
2. BB2 = size
 Hover name
15 μL/lane; 1% agarose; Ladder
DNA(plasmid) 3.0 μL
10X buffer 1.5
EcoRI 1.0
PstI 1.0
dH2O 8.5
  15 μL --> 37°C/ ~15 min.



Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Karmella_Haynes/Notebook/BioBrick_cloning/2013/04/01&oldid=687976"