User:Karmella Haynes/Notebook/PcTF Genomics/2014/02/14

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(02/14/14)
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* Resuspended each pellet in 600 μL dH<sub>2</sub>O before miniprep
* Resuspended each pellet in 600 μL dH<sub>2</sub>O before miniprep
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{| class="wikitable" border="0" cellspacing="3" <!-- Rxn. table -->
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{| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table -->
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|-valign="top"
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|- valign="top"
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| <u>Reagent</u> || <u>Volume</u>  
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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. CMV = 3200, 700<br>2. CMV = 3200, 700<br>3. MV9 = 3200, 700<br>4. MV9 = 3200, 700<br>
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| rowspan="7" | <!-- [[Image:GelImage.jpg|400px|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] -->
|-
|-
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| reagent 1 || # μL
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| DNA(plasmid) || 3.0 μL
|-
|-
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| reagent 2 || #
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| 10X buffer || 1.5
|-
|-
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| reagent 3 || #
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| PstI || 1.0
|-
|-
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| reagent 4 || #
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| EcoRI or XbaI || 1.0
|-
|-
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| dH<sub>2</sub>O || #
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| dH<sub>2</sub>O || 8.5
|-
|-
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| &nbsp; || # μL
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| &nbsp; || 15 μL --> 37°C/ ~10 min.
|}
|}
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--> Reaction conditions
--> Reaction conditions

Revision as of 18:38, 14 February 2014

Pc-TF Genomics Main project page
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02/14/14

  • Miniprep digests: CMV/V0120, MV9

Miniprep Digests

  1. CMV culture 1 - check with E/P
  2. CMV culture 2 - check with E/P
  3. MV9 culture 1 - check with P/X
  4. MV9 culture 2 - check with P/X
  • Used the Zymo miniprep kit
  • Pelleted each 5 mL culture at 2000 rpm/ 5 min in old centrifuge
  • Resuspended each pellet in 600 μL dH2O before miniprep
Reagent Volume Expected:
1. CMV = 3200, 700
2. CMV = 3200, 700
3. MV9 = 3200, 700
4. MV9 = 3200, 700
DNA(plasmid) 3.0 μL
10X buffer 1.5
PstI 1.0
EcoRI or XbaI 1.0
dH2O 8.5
  15 μL --> 37°C/ ~10 min.


--> Reaction conditions



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