User:Wesley J. Houston/Notebook/MAP/2013/03/13

From OpenWetWare
< User:Wesley J. Houston‎ | Notebook‎ | MAP‎ | 2013‎ | 03
Revision as of 22:12, 25 March 2013 by Wesley J. Houston (talk | contribs) (3/13/13)
Jump to: navigation, search
Owwnotebook icon.png 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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

3/13/13

  • Attempted ligation following the same protocol outlined on 3/4/13.
  • Grew four liquid cultures of KAH182, following the same procedure (pick a colony and place into LB broth), in case this ligation attempt fails.

> Ligations

  1 2
Insert DNA (KAH182) 3.0 ---
Vector DNA (KAH201) 0.5 0.5
2x lgn buf (Roche) 5.0 5.0
T4 ligase (NEB) 1.0 1.0
dH2O 0.5 3.5
  10 μL 10 μL
  • Follow quick transformation protocol
  • Add 30 uL DH5α-Turbo to each ligation; plate on 100 μg/mL Amp agar

> Ligations (corrected version, 3/04/13)

Ligation Plate results (lig : neg crtl) 03/13/13
1. KAH201 (S/P)/3403, 25 ng + KAH182 (X/P)/1675, 26 ng 0
2. KAH201 (S/P)/3403, 25 ng 0