User:Karmella Haynes/Notebook/PcTF Genomics/2014/02/14: Difference between revisions

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==mm/dd/yy==
==02/14/14==
<!-- Precede finished items with a checkmark &#x2713; -->
<!-- Precede finished items with a checkmark &#x2713; -->
* Line item 1
* Miniprep digests: CMV/V0120, MV9
 


----
----
'''Line item 1'''<br>
'''Miniprep Digests'''<br>
> Samples
# CMV culture 1 - check with E/P
# CMV culture 2 - check with E/P
# MV9 culture 1 - check with P/X
# 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 dH<sub>2</sub>O before miniprep


{| class="wikitable" border="0" cellspacing="3" <!-- Rxn. table -->
{| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table -->
|-valign="top"
|- valign="top"
| <u>Reagent</u> || <u>Volume</u>
| bgcolor=#cfcfcf | Reagent
| bgcolor=#cfcfcf | Volume
| rowspan="7" | <u>Expected (approx):</u><br>1. CMV = 3200, 700<br>2. CMV = 3200, 700<br>3. MV9 = 4550, 60<br>4. MV9 = 4550, 60<br>
| rowspan="7" | [[Image:KAH021414_gel1.jpg|400px|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
|-
|-
| reagent 1 || # μL
| DNA(plasmid) || 3.0 μL
|-
|-
| reagent 2 || #
| 10X buffer || 1.5
|-
|-
| reagent 3 || #
| PstI || 1.0
|-
|-
| reagent 4 || #
| EcoRI or XbaI || 1.0
|-
|-
| dH<sub>2</sub>O || #
| dH<sub>2</sub>O || 8.5
|-
|-
| &nbsp; || # μL
| &nbsp; || 15 μL --> 37°C/ ~10 min.
|}
|}


--> Reaction conditions
 
* Gave these to Cameron Gardner to build CMV/MV9
 




Revision as of 12:37, 2 April 2014

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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 (approx):
1. CMV = 3200, 700
2. CMV = 3200, 700
3. MV9 = 4550, 60
4. MV9 = 4550, 60
 Hover name
15 μL/lane; 1% agarose; Ladder
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.


  • Gave these to Cameron Gardner to build CMV/MV9




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