User:Wesley J. Houston/Notebook/MAP/2013/03/13

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(3/13/13)
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==3/13/13==
==3/13/13==
-
* Attempt ligation again with original plasmids.
+
* Attempted ligation following the same protocol outlined on 3/4/13.
-
* Grow liquid cultures of KAH182 in case this ligation attempt fails.
+
* Grew four liquid cultures of KAH182, following the same procedure (pick a colony and place into LB broth), in case this ligation attempt fails.
 +
{| class="wikitable" border="0" cellspacing="3" <!-- Ligation rxn table -->
 +
| &nbsp;            || 1    || 2    ||
 +
|-
 +
| Insert DNA (KAH182)        || 3.0  || ---  ||
 +
|-
 +
| Vector DNA (KAH201)        || 0.5  || 0.5  ||
 +
|-
 +
| 2x lgn buf (Roche) || 5.0  || 5.0  ||
 +
|-
 +
| T4 ligase (NEB)    || 1.0  || 1.0  ||
 +
|-
 +
| dH<sub>2</sub>O    ||  0.5 ||  3.5 ||
 +
|-
 +
| &nbsp;            || 10 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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Revision as of 16:07, 18 March 2013

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3/13/13

  • Attempted ligation following the same protocol outlined on 3/4/13.
  • Grew four liquid cultures of KAH182, following the same procedure (pick a colony and place into LB broth), in case this ligation attempt fails.
  1 2
Insert DNA (KAH182) 3.0 ---
Vector DNA (KAH201) 0.5 0.5
2x lgn buf (Roche) 5.0 5.0
T4 ligase (NEB) 1.0 1.0
dH2O 0.5 3.5
  10 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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