User:Wesley J. Houston/Notebook/MAP/2013/03/11

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(3/11/13)
Line 8: Line 8:
==3/11/13==
==3/11/13==
* Grow liquid cultures of KAH182 to repeat ligation.
* Grow liquid cultures of KAH182 to repeat ligation.
-
* Retry ligation with lower concentration plasmids (will make table).
+
* Retry ligation with lower concentration plasmids.
 +
 
 +
{| class="wikitable" border="0" cellspacing="3" <!-- Ligation rxn table -->
 +
| &nbsp;            || 1    || 2    ||
 +
|-
 +
| Insert DNA (KAH182)       || 4.0  || ---  ||
 +
|-
 +
| Vector DNA (KAH201)        || 0.65  || 0.65  ||
 +
|-
 +
| 2x lgn buf (Roche) || 5.0  || 5.0  ||
 +
|-
 +
| T4 ligase (NEB)    || 1.0  || 1.0  ||
 +
|-
 +
| dH<sub>2</sub>O    ||  0 ||  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
 +
 

Revision as of 16:12, 18 March 2013

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3/11/13

  • Grow liquid cultures of KAH182 to repeat ligation.
  • Retry ligation with lower concentration plasmids.
  1 2
Insert DNA (KAH182) 4.0 ---
Vector DNA (KAH201) 0.65 0.65
2x lgn buf (Roche) 5.0 5.0
T4 ligase (NEB) 1.0 1.0
dH2O 0 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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