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

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| bgcolor=#cfcfcf | Reagent
| bgcolor=#cfcfcf | Reagent
| bgcolor=#cfcfcf | Volume  
| bgcolor=#cfcfcf | Volume  
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| rowspan="7" | <u>Expected:</u><br>1. H2B = 378<br>2. LOV = 450<br>3, 4. pSB1A3 = ~2000
+
| rowspan="7" | <u>Expected:</u><br>1. KAH109 = 1132 <br>2. KAH111 = 1132 <br>3. KAH112 = 1132 <br>4. KAH115 = 931 <br>5. KAH148 = 967 <br> 6. KAH150 = 967 <br> 7. KAH151 = 967
| 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]
| 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]
|-
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Revision as of 18:39, 5 March 2013

PcTF Subcloning in E. coli Main project page
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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. KAH109 = 1132
2. KAH111 = 1132
3. KAH112 = 1132
4. KAH115 = 931
5. KAH148 = 967
6. KAH150 = 967
7. KAH151 = 967 		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]
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