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

< User:Wesley J. Houston | Notebook | MAP | 2013 | 03(Difference between revisions)

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Project name

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.

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-==Entry title==
+==3/18/2013==
-* Insert content here...
+* 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
+{| 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>
+|-
+| 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 || 0
+|}
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