User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/10/31: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
(Autocreate 2013/10/31 Entry for User:Behzad_Damadzadeh/Notebook/PcTF_Subcloning_in_E-coli)
 
Line 7: Line 7:
<!-- ##### 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. ##### -->


=Date=
=== Site Directed Mutagenesis of BD002 Ligation (Second Try) ===


'''List title'''
 
# List items
*'''Ligate The Sticky Ends to Make Circular DNA'''
 
{| {{table}} cellspacing="7"  width=650px
| Sample type || colspan="3"  style="text-align: center;" |  Tube 1 || colspan="3"  style="text-align: center;" |  Tube 2 ||  style="text-align: center;" |  (-) Ctrl
|-
| DNA Conc. (ng) || 10ng || 30ng || 50ng || 10ng || 30ng || 50ng ||  ---
|-
| DNA μL || 1μL || 3μL || 5μL || 1μL || 3μL || 5μL || 5μL
|-
| T4 Ligase  || 1.0 μl || 1.0μL || 1.0μL || 1.0μL || 1.0μL || 1.0μL || 0.0μL
|-
| 2X Roche Buffer|| 5.00 μl || 5.00 μL || 6.00 μL || 5.00 μL || 5.00 μL || 6.00 μL ||  5.00 μL
|-
| dH<sub>2</sub>O || 3.00 μl || 1.00 μl || ---  || 3.00 μL || 1.00 μL || --- || ---
|-
| Total || 10.0 μl || 10.0 μl || 12.0 μl || 10.0 μl || 10.0 μl || 12.00 μL || 10.00 μL 
|-
| Incubate at room temperature for 30 minutes.
|-
| '''Transformation result''' || No colony || No colony || No colony || <span style="color:#FF0000">Two Colonies</span>|| <span style="color:#FF0000">No Colony</span>
|}
 
* Fast transformation protocol in DH5α-T. Add 30 μL of cells to each sample and incubate on ice for 30 minutes.
* Grow on AMP agar plates and incubate overnight at 37°C.

Revision as of 16:52, 31 October 2013

PcTF Subcloning in E. coli <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>

Site Directed Mutagenesis of BD002 Ligation (Second Try)

  • Ligate The Sticky Ends to Make Circular DNA
Sample type Tube 1 Tube 2 (-) Ctrl
DNA Conc. (ng) 10ng 30ng 50ng 10ng 30ng 50ng ---
DNA μL 1μL 3μL 5μL 1μL 3μL 5μL 5μL
T4 Ligase 1.0 μl 1.0μL 1.0μL 1.0μL 1.0μL 1.0μL 0.0μL
2X Roche Buffer 5.00 μl 5.00 μL 6.00 μL 5.00 μL 5.00 μL 6.00 μL 5.00 μL
dH2O 3.00 μl 1.00 μl --- 3.00 μL 1.00 μL --- ---
Total 10.0 μl 10.0 μl 12.0 μl 10.0 μl 10.0 μl 12.00 μL 10.00 μL
Incubate at room temperature for 30 minutes.
Transformation result No colony No colony No colony Two Colonies No Colony
  • Fast transformation protocol in DH5α-T. Add 30 μL of cells to each sample and incubate on ice for 30 minutes.
  • Grow on AMP agar plates and incubate overnight at 37°C.
Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Behzad_Damadzadeh/Notebook/PcTF_Subcloning_in_E-coli/2013/10/31&oldid=753227"