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

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Line 10: Line 10:
* Retry ligation with lower concentration plasmids.
* Retry ligation with lower concentration plasmids.


> Ligations
{| class="wikitable" border="0" cellspacing="3" <!-- Ligation rxn table -->
{| class="wikitable" border="0" cellspacing="3" <!-- Ligation rxn table -->
| &nbsp;            || 1    || 2    ||
| &nbsp;            || 1    || 2    ||
Line 28: Line 29:
* Follow quick transformation protocol
* Follow quick transformation protocol
* Add 30 uL DH5α-Turbo to each ligation; plate on 100 μg/mL Amp agar
* 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> 03/11/13</font>
|-
| 1. KAH201 (S/P)/<font color="blue">3403</font>, 25 ng + KAH182 (X/P)/1675, 26 ng || <font color="blue">10</font>
|-
| 2. KAH201 (S/P)/3403, 25 ng || 0
|}




Revision as of 22:11, 25 March 2013

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3/11/13

  • Grow liquid cultures of KAH182 to repeat ligation.
  • Retry ligation with lower concentration plasmids.

> Ligations

  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
Ligation Plate results (lig : neg crtl) 03/11/13
1. KAH201 (S/P)/3403, 25 ng + KAH182 (X/P)/1675, 26 ng 10
2. KAH201 (S/P)/3403, 25 ng 0



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