Haynes Lab:Notebook/Investigating Photo-Switchable Synthetic Nucleosomes/2013/03/13: Difference between revisions
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# Roche - pSB1A3 | # Roche - pSB1A3 | ||
{| {{table}} border="1" cellspacing="3" <!-- Golden Gate Rxn. table --> | {| {{table}} border="1" cellspacing="3" <!-- Golden Gate Rxn. table --> | ||
|- valign="top" | |- valign="top" | ||
| Reagent || Promega (1 | | Reagent || Promega (1) || Promega (2) || Roche (9, 10) || Roche (11, 12) | ||
|- | |- | ||
| gg2 pSB1A3* | | gg2 pSB1A3* || 1.0 || 1.0 || 1.0 || 1.0 | ||
|- | |- | ||
| gg3 hPCD* | | gg3 hPCD* || 1.0 || --- || 1.0 || --- | ||
|- | |- | ||
| gg4 BL01* | | gg4 BL01* || 1.0 || --- || 1.0 || --- | ||
|- | |- | ||
| ligase buffer | | ligase buffer || 1.0 || 1.0|| 5.0 || 5.0 | ||
|- | |- | ||
| NEB T4 ligase | | NEB T4 ligase || 0.25 || 0.25 || 0.25 || 0.25 | ||
|- | |- | ||
| NEB BsmBI | | NEB BsmBI || 0.5 || 0.5 || 0.5 || 0.5 | ||
|- | |- | ||
| dH<sub>2</sub>O | | dH<sub>2</sub>O || 5.25 || 7.25 || 1.25 || 3.25 | ||
|- | |- | ||
| | | || 10.0 || 10.0 || 10.0 || 10.0 | ||
|} | |} | ||
Note: *DNA is 20 fmole/μL | Note: *DNA is 20 fmole/μL | ||
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* Transformations | * Transformations | ||
** | ** 50 μL '''BL21''' in 2.0 mL tubes; ice 2 min.; 42°C 90 sec.; add 800 μL SOC medium; shake @ 37°C 25 min.; pellet @ top speed 3 min.; resuspend in 100 μL amp liq. medium; plate on amp agar | ||
Revision as of 21:31, 12 March 2013
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03/13/13
Golden Gate Assembly
Note: *DNA is 20 fmole/μL Thermal cycler
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