Haynes Lab:Notebook/Engineering PC-TFs/2014/04/18

From OpenWetWare
Jump to navigationJump to search


<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>

Summary

4/16

  • Cloned mv9cmv construct into DH5a-T cells using standard procedure. Incubated overnight.

4/17

  • Removed plates from incubator.
Hover name;
Hover name
;
  • 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.
Plasmid OD260 OD260/280 ng/μL
1. Colony 1 0.202 1.905 162
2. Colony 2 0.253 1.92 203
  • Restriction Digest
Reagent Volume Expected:
1. CMV = 643bp
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.
Hover name
15 μL/lane; 1% agarose; 
Ladder
  • Expected base pair length: 643
Retrieved from "https://openwetware.org/mediawiki/index.php?title=Haynes_Lab:Notebook/Engineering_PC-TFs/2014/04/18&oldid=789376"