User:Wesley J. Houston/Notebook/MAP/2013/04/01: Difference between revisions
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(Autocreate 2013/04/01 Entry for User:Wesley_J._Houston/Notebook/MAP) |
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== | ==4/1/13== | ||
* | * Completed 2 ligations of KAH201 (S/P)/3403 + KAH182 (X/P)/1675 | ||
** First ligation used KAH201 (S/P)/3403 and KAH182 (X/P) 1675 made on [http://openwetware.org/wiki/User:Wesley_J._Houston/Notebook/MAP/2013/03/26 3/26/13] | |||
** Second ligation used the same KAH182 (X/P) 1675 and KAH201 (S/P)/3403 made on [http://openwetware.org/wiki/User:Wesley_J._Houston/Notebook/MAP/2013/02/08 2/8/13] | |||
> Ligation 1 | |||
{| class="wikitable" border="0" cellspacing="3" <!-- Ligation rxn table --> | |||
| || 1 || 2 || | |||
|- | |||
| Insert DNA (KAH182) || 3.5 || --- || | |||
|- | |||
| Vector DNA (KAH201) || 1.0 || 1.0 || | |||
|- | |||
| 2x lgn buf (Roche) || 5.5 || 5.5 || | |||
|- | |||
| T4 ligase (NEB) || 1.0 || 1.0 || | |||
|- | |||
| dH<sub>2</sub>O || 0.0 || 3.5 || | |||
|- | |||
| || 11 μL || 11 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 1</font> | |||
|- | |||
| 1. KAH201 (S/P)/<font color="blue">3403</font>, 25 ng + KAH182 (X/P)/1675, 26 ng || <font color="blue">0</font> | |||
|- | |||
| 2. KAH201 (S/P)/3403, 25 ng || 0 | |||
|} | |||
> Ligation 2 | |||
{| class="wikitable" border="0" cellspacing="3" <!-- Ligation rxn table --> | |||
| || 1 || 2 || | |||
|- | |||
| Insert DNA (KAH182) || 3.5 || --- || | |||
|- | |||
| Vector DNA (KAH201) || 0.7 || 0.7 || | |||
|- | |||
| 2x lgn buf (Roche) || 5.2 || 5.2 || | |||
|- | |||
| T4 ligase (NEB) || 1.0 || 1.0 || | |||
|- | |||
| dH<sub>2</sub>O || 0.6 || 4.1 || | |||
|- | |||
| || 11 μL || 11 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 2</font> | |||
|- | |||
| 1. KAH201 (S/P)/<font color="blue">3403</font>, 25 ng + KAH182 (X/P)/1675, 26 ng || <font color="blue">~40</font> | |||
|- | |||
| 2. KAH201 (S/P)/3403, 25 ng || 0 | |||
|} | |||
* Ligation 2 appears successful, with ~40 colonies in the ligation plate and 0 colonies in the negative control plate. [[Image:4.1.13ligation.jpg |150px|Results of the ligation]] | |||
** The next step will be to confirm the assembly, to be completed on [http://openwetware.org/wiki/User:Wesley_J._Houston/Notebook/MAP/2013/04/04 4/4/13] | |||
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Revision as of 18:19, 2 April 2013
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4/1/13
> Ligation 1
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