Difference between revisions of "User:Wesley J. Houston/Notebook/MAP/2013/03/18"

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* Retrying ligation of KAH201(S/P)/3403 + KAH182(X/P)/1675
 
* Retrying ligation of KAH201(S/P)/3403 + KAH182(X/P)/1675
  
> Ligations, following corrected version from [http://openwetware.org/wiki/User:Wesley_J._Houston/Notebook/MAP/2013/02/08| March 4, 2013], with 50 ng of vector, instead of 25 ng.
+
> Ligations, following corrected version from [http://openwetware.org/wiki/User:Wesley_J._Houston/Notebook/MAP/2013/02/08|March 4, 2013], with 50 ng of vector, instead of 25 ng.
 
{| class="wikitable" border="0" cellspacing="3" <!-- Ligations table -->
 
{| class="wikitable" border="0" cellspacing="3" <!-- Ligations table -->
 
| <u>Ligation</u> || <font color="blue"><u>Plate results (lig : neg crtl)</u> 03/18/13</font>
 
| <u>Ligation</u> || <font color="blue"><u>Plate results (lig : neg crtl)</u> 03/18/13</font>

Revision as of 11:10, 18 March 2013

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3/18/2013

  • Retrying ligation of KAH201(S/P)/3403 + KAH182(X/P)/1675

> Ligations, following corrected version from 4, 2013, with 50 ng of vector, instead of 25 ng.

Ligation Plate results (lig : neg crtl) 03/18/13
1. KAH201 (S/P)/3403, 50 ng + KAH182 (X/P)/1675, 50 ng TBD
2. KAH201 (S/P)/3403, 50 ng TBD
  1 2
Insert DNA (KAH182) 6.0 ---
Vector DNA (KAH201) 1.0 1.0
2x lgn buf (Roche) 5.0 5.0
T4 ligase (NEB) 1.0 1.0
dH2O 0 3.0
  13 μL 10 μL
  • Follow quick transformation protocol
  • Add 30 uL DH5α-Turbo to each ligation; plate on 100 μg/mL Amp agar