User:Wesley J. Houston/Notebook/MAP/2013/04/04: Difference between revisions

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(Autocreate 2013/04/04 Entry for User:Wesley_J._Houston/Notebook/MAP)
 
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==Entry title==
==4/4/13==
* Insert content here...
* The liquid cultures did not grow.
* Prepped two more liquid cultures from the ligation plate, as well as performed another ligation.


> Ligation
{| class="wikitable" border="0" cellspacing="3" <!-- Ligation rxn table -->
| &nbsp;            || 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  ||
|-
| dH<sub>2</sub>O    ||  0.6 ||  7.6 ||
|-
| &nbsp;            || 20 μL || 20 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>04/01/13: Ligation</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
|}
* The ligation was unsuccessful.


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Revision as of 22:27, 7 April 2013

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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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