User:Karmella Haynes/Notebook/BioBrick cloning/2013/02/23: Difference between revisions
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(02/24/13)<br> | |||
Golden Gate Reactions | Golden Gate Reactions | ||
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{| {{table}} border="1" cellspacing="3" <!-- Golden Gate Rxn. table --> | {| {{table}} border="1" cellspacing="3" <!-- Golden Gate Rxn. table --> | ||
|- valign="top" | |- valign="top" | ||
| Reagent || 1 || 2 | | Reagent || 1 || 2 || Master mix (x3) | ||
|- | |- | ||
| gg2 pSB1A3 || 1.0 || 1.0 | | gg2 pSB1A3 || 1.0 || 1.0 || --- | ||
|- | |- | ||
| gg3 | | gg3 LOV || 1.0 || H2O || --- | ||
|- | |- | ||
| gg4 | | gg4 H2B+his || 1.0 || H2O || --- | ||
|- | |- | ||
| 10x PRO ligase buffer || 1.0 || 1.0 | | 10x PRO ligase buffer || 1.0 || 1.0 || 3.0 | ||
|- | |- | ||
| NEB T4 lgase || 0.25 || 0.25 | | NEB T4 lgase || 0.25 || 0.25 || 0.75 | ||
|- | |- | ||
| BsmBI || 0.5 || 0.5 | | BsmBI || 0.5 || 0.5 || 1.5 | ||
|- | |- | ||
| dH<sub>2</sub>O || 5.25 || | | dH<sub>2</sub>O || 5.25 || 5.25 || 15.75 | ||
|- | |- | ||
| || 10.0 || 10.0 | | || 10.0 || 10.0 | ||
|} | |} | ||
* Aliquot 7.0 mix to each tube; add DNA/ H2O | |||
Thermal cycling | Thermal cycling |
Revision as of 18:34, 23 February 2013
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02/23/13
Type IIS Assembly, PCR: LOV-H2B+his
PCR
Golden Gate Reactions
Thermal cycling
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