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

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(Autocreate 2013/03/05 Entry for User:Behzad_Damadzadeh/Notebook/PcTF_Subcloning_in_E-coli)
Current revision (19:19, 5 March 2013) (view source)
(3/5/2013)
 
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=Date=
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=3/5/2013=
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'''List title'''
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'''PcTF + His Tag Ligation in V0120'''
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# List items
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PCR
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* 95°C, 3 min.
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* [95°C, 30 sec.; 57°C, 30 sec.; 72°C, 1 min.] x35
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* 72°C, 3 min.
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* 4°C, ∞
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{| {{table}} border="1" cellspacing="3" <!-- PCR rxn. table -->
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|- valign="top"
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| bgcolor=#cfcfcf | Reagent
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| bgcolor=#cfcfcf | Volume
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| 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
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| rowspan="7" | [[Image:PCR KAH Product.png|400px|Hover name]]<br>10 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
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|-
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| DNA(plasmid) || 0.2 μL
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|-
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| 10 μM primer 1 || 1.0
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|-
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| 10 μM primer 2 || 1.0
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|-
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| 2x GoTaq || 25.0
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|-
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| dH<sub>2</sub>O || 22.8
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|-
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| &nbsp; || 50.0 μL
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|}
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Purification
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* Clean with Zymo DNA c&c; elute w/ 20 μL dH<sub>2</sub>O
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* Measure [DNA]

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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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