User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/03/05

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(Autocreate 2013/03/05 Entry for User:Behzad_Damadzadeh/Notebook/PcTF_Subcloning_in_E-coli)
(Date)
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=
+
=3/5/2013=
-
'''List title'''
+
'''PcTF + His Tag Ligation in V0120'''
-
# List items
+
 
 +
PCR
 +
* 95°C, 3 min.
 +
* [95°C, 30 sec.; 57°C, 30 sec.; 72°C, 1 min.] x35
 +
* 72°C, 3 min.
 +
* 4°C, ∞
 +
 
 +
{| {{table}} border="1" cellspacing="3" <!-- PCR rxn. table -->
 +
|- valign="top"
 +
| bgcolor=#cfcfcf | Reagent
 +
| bgcolor=#cfcfcf | Volume
 +
| rowspan="7" | <u>Expected:</u><br>1. H2B = 378<br>2. LOV = 450<br>3, 4. pSB1A3 = ~2000
 +
| rowspan="7" | [[Image:KAH022313_gel1.jpg|200px|Hover name]]<br>10 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
 +
|-
 +
| DNA(plasmid) || 0.2 μL
 +
|-
 +
| 10 μM primer 1 || 1.0
 +
|-
 +
| 10 μM primer 2 || 1.0
 +
|-
 +
| 2x GoTaq || 25.0
 +
|-
 +
| dH<sub>2</sub>O || 22.8
 +
|-
 +
| &nbsp; || 50.0 μL
 +
|}
 +
 
 +
 
 +
Purification
 +
* Clean with Zymo DNA c&c; elute w/ 20 μL dH<sub>2</sub>O
 +
* Measure [DNA]

Revision as of 18:35, 5 March 2013

PcTF Subcloning in E. coli Main project page
Previous entry      Next entry

3/5/2013

PcTF + His Tag Ligation in V0120

PCR

  • 95°C, 3 min.
  • [95°C, 30 sec.; 57°C, 30 sec.; 72°C, 1 min.] x35
  • 72°C, 3 min.
  • 4°C, ∞
Reagent Volume Expected:
1. H2B = 378
2. LOV = 450
3, 4. pSB1A3 = ~2000
Hover name
10 μL/lane; 1% agarose; Ladder
DNA(plasmid) 0.2 μL
10 μM primer 1 1.0
10 μM primer 2 1.0
2x GoTaq 25.0
dH2O 22.8
  50.0 μL


Purification

  • Clean with Zymo DNA c&c; elute w/ 20 μL dH2O
  • Measure [DNA]
Personal tools