User:Karmella Haynes/Notebook/BioBrick cloning/2012/12/03: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
(One intermediate revision by the same user not shown)
Line 6: Line 6:
| colspan="2"|
| colspan="2"|
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
==mm/dd/yy==
==12/03/12==
<!-- Precede finished items with a checkmark &#x2713; -->
<!-- Precede finished items with a checkmark &#x2713; -->
* Assembly: KAH111 + C-His expression vector (trouble shooting for Behzad)
* Assembly: KAH111 + C-His expression vector (trouble shooting for Behzad)
Line 37: Line 37:


{| {{table}} cellspacing="3" <!-- Ligation rxn table -->
{| {{table}} cellspacing="3" <!-- Ligation rxn table -->
| &nbsp;            || 1    || 2    ||
| &nbsp;            || 1    || 2     
|-
|-
| Insert DNA        || 1.0  || ---  ||
| Insert DNA        || 1.0  || ---   
|-
|-
| Vector DNA        || 0.5  || 0.5  ||
| Vector DNA        || 0.5  || 0.5   
|-
|-
| 2x lgn buf (Roche) || 5.0  || 5.0  ||
| 2x lgn buf (Roche) || 5.0  || 5.0   
|-
|-
| T4 ligase (NEB)    || 1.0  || 1.0  ||
| T4 ligase (NEB)    || 1.0  || 1.0   
|-
|-
| dH<sub>2</sub>O    || 2.5  || 3.5  ||
| dH<sub>2</sub>O    || 2.5  || 3.5   
|-
|-
| &nbsp;            || 10 μL || 10 μL ||
| &nbsp;            || 10 μL || 10 μL  
|}
|}


Revision as of 19:40, 3 December 2012

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>

12/03/12

  • Assembly: KAH111 + C-His expression vector (trouble shooting for Behzad)


Assemblies

  1. KAH111/C-His: KAH111/(E/P)/1123 + C-His/(E/P)/3600


  • Digests (Fermentas FD)
    • Already done by Behzad


  • Measure conc.'s
    • Behzad: KAH111 = 15 ng/μL
    • Behzad: C-His = 57 ng/μL


  • Ligations
Ligation Plate results (lig : neg crtl) mm/dd/yy
1. KAH111(E/P)/1123, 16 ng + C-His(E/P)/3600, 25 ng new BioBrick #:1 (Pick #)
2. C-His(E/P)/ 25 ng  
  1 2
Insert DNA 1.0 ---
Vector DNA 0.5 0.5
2x lgn buf (Roche) 5.0 5.0
T4 ligase (NEB) 1.0 1.0
dH2O 2.5 3.5
  10 μL 10 μL



Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Karmella_Haynes/Notebook/BioBrick_cloning/2012/12/03&oldid=662121"