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

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==3/18/2013==
 
==3/18/2013==
 
* Retrying ligation of KAH201(S/P)/3403 + KAH182(X/P)/1675
 
* Retrying ligation of KAH201(S/P)/3403 + KAH182(X/P)/1675
 +
* Concentrations: KAH201 = 48.1 ng/uL, KAH1182 = 6.3 ng/uL
 +
** Concentrations were determined on 2/18/13
 +
 +
> Ligations, following corrected version from 3/4/13, with 50 ng of vector instead of 25 ng.
 +
{| class="wikitable" border="0" cellspacing="3" <!-- Ligation rxn table -->
 +
| &nbsp;            || 1    || 2    ||
 +
|-
 +
| Insert DNA (KAH182)|| 8.0  || ---  ||
 +
|-
 +
| Vector DNA (KAH201)|| 1.0  || 1.0  ||
 +
|-
 +
| 2x lgn buf (Roche) || 10.0  || 5.0  ||
 +
|-
 +
| T4 ligase (NEB)    || 1.0  || 1.0  ||
 +
|-
 +
| dH<sub>2</sub>O    ||  0  || 3.0  ||
 +
|-
 +
| &nbsp;            || 20 μL || 10 μL ||
 +
|}
 +
 +
* Follow quick transformation protocol
 +
* Add 30 uL DH5α-Turbo to each ligation; plate on 100 μg/mL Amp agar
  
> Ligations (corrected version, 3/04/13)
 
 
{| 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>
 
|-
 
|-
| 1. KAH201 (S/P)/<font color="blue">3403</font>, 50 ng + KAH182 (X/P)/1675, 50 ng || <font color="blue">TBD</font>
+
| 1. KAH201 (S/P)/<font color="blue">3403</font>, 50 ng + KAH182 (X/P)/1675, 50 ng || <font color="blue">0</font>
 
|-
 
|-
| 2. KAH201 (S/P)/3403, 50 ng || TBD
+
| 2. KAH201 (S/P)/3403, 50 ng || 0
 
|}
 
|}
 
  
 
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Revision as of 21:13, 25 March 2013

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3/18/2013

  • Retrying ligation of KAH201(S/P)/3403 + KAH182(X/P)/1675
  • Concentrations: KAH201 = 48.1 ng/uL, KAH1182 = 6.3 ng/uL
    • Concentrations were determined on 2/18/13

> Ligations, following corrected version from 3/4/13, with 50 ng of vector instead of 25 ng.

  1 2
Insert DNA (KAH182) 8.0 ---
Vector DNA (KAH201) 1.0 1.0
2x lgn buf (Roche) 10.0 5.0
T4 ligase (NEB) 1.0 1.0
dH2O 0 3.0
  20 μL 10 μL
  • Follow quick transformation protocol
  • Add 30 uL DH5α-Turbo to each ligation; plate on 100 μg/mL Amp agar
Ligation Plate results (lig : neg crtl) 03/18/13
1. KAH201 (S/P)/3403, 50 ng + KAH182 (X/P)/1675, 50 ng 0
2. KAH201 (S/P)/3403, 50 ng 0