User:Wesley J. Houston/Notebook/MAP/2013/03/18

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(3/18/2013)
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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
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> Ligations (corrected version, 3/04/13)
+
> Ligations, following corrected version from [[3/4/13|http://openwetware.org/wiki/User:Wesley_J._Houston/Notebook/MAP/2013/02/08]], 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>
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| 2. KAH201 (S/P)/3403, 50 ng || TBD
| 2. KAH201 (S/P)/3403, 50 ng || TBD
|}
|}
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 +
{| class="wikitable" border="0" cellspacing="3" <!-- Ligation rxn table -->
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| &nbsp;            || 1    || 2    ||
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|-
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| Insert DNA (KAH182)|| 6.0  || ---  ||
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|-
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| Vector DNA (KAH201)|| 1.0  || 1.0  ||
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|-
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| 2x lgn buf (Roche) || 5.0  || 5.0  ||
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|-
 +
| T4 ligase (NEB)    || 1.0  || 1.0  ||
 +
|-
 +
| dH<sub>2</sub>O    ||  0  || 3.0  ||
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|-
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| &nbsp;            || 13 μL || 10 μL ||
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|}
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* Follow quick transformation protocol
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* Add 30 uL DH5α-Turbo to each ligation; plate on 100 μg/mL Amp agar
 +

Revision as of 14:07, 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 http://openwetware.org/wiki/User:Wesley_J._Houston/Notebook/MAP/2013/02/08, 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



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