User:Wesley J. Houston/Notebook/MAP/2013/04/04

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(Autocreate 2013/04/04 Entry for User:Wesley_J._Houston/Notebook/MAP)
Current revision (00:27, 8 April 2013) (view source)
(Entry title)
 
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==Entry title==
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==4/4/13==
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* Insert content here...
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* The liquid cultures did not grow.
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* Prepped two more liquid cultures from the ligation plate, as well as performed another ligation.
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> Ligation
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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)        || 7.0  || ---  ||
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|-
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| Vector DNA (KAH201)        || 1.7  || 1.7  ||
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|-
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| 2x lgn buf (Roche) || 9.7  || 9.7  ||
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|-
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| T4 ligase (NEB)    || 1.0  || 1.0  ||
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|-
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| dH<sub>2</sub>O    ||  0.6 ||  7.6 ||
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|-
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| &nbsp;            || 20 μL || 20 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
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{| class="wikitable" border="0" cellspacing="3" <!-- Ligations table -->
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| <u>Ligation</u> || <font color="blue"><u>Plate results (lig : neg crtl)</u>04/01/13: Ligation</font>
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|-
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| 1. KAH201 (S/P)/<font color="blue">3403</font>, 50 ng + KAH182 (X/P)/1675, 50 ng || <font color="blue">0</font>
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|-
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| 2. KAH201 (S/P)/3403, 50 ng || 0
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|}
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* The ligation was unsuccessful.
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4/4/13

  • The liquid cultures did not grow.
  • Prepped two more liquid cultures from the ligation plate, as well as performed another ligation.

> Ligation

  1 2
Insert DNA (KAH182) 7.0 ---
Vector DNA (KAH201) 1.7 1.7
2x lgn buf (Roche) 9.7 9.7
T4 ligase (NEB) 1.0 1.0
dH2O 0.6 7.6
  20 μL 20 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)04/01/13: Ligation
1. KAH201 (S/P)/3403, 50 ng + KAH182 (X/P)/1675, 50 ng 0
2. KAH201 (S/P)/3403, 50 ng 0
  • The ligation was unsuccessful.


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