User:Karmella Haynes/Notebook/BioBrick cloning/2013/01/15: Difference between revisions
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* Make Promega buffer from scratch. | * Make Promega buffer from scratch. | ||
* | |||
* | |||
* Golden Gate assembly reactions | |||
1, 2. Promega - hPCD+BL01+pSB1A3<br> | |||
3, 4. Promega - pSB1A3<br> | |||
5, 6. NEB - hPCD+BL01+pSB1A3<br> | |||
7, 8. NEB - pSB1A3<br> | |||
9, 10. Roche - hPCD+BL01+pSB1A3<br> | |||
11, 12. Roche - pSB1A3<br> | |||
{| {{table}} border="1" cellspacing="3" <!-- Golden Gate Rxn. table --> | |||
|- valign="top" | |||
| Reagent || Promega (1,2) || Promega (3, 4) || NEB (5, 6) || NEB (7, 8) || Roche (9, 10) || Roche (11, 12) | |||
|- | |||
| gg2 pSB1A3 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 || 1.0 | |||
|- | |||
| gg3 hPCD || 1.0 || --- || 1.0 || --- || 1.0 || --- | |||
|- | |||
| gg4 BL01 || 1.0 || --- | |||
|- | |||
| 10x ligase buffer || 1.0 || 1.0 | |||
|- | |||
| NEB T4 lgase || 0.25 || 0.25 | |||
|- | |||
| BsmBI || 0.5 || 0.5 | |||
|- | |||
| dH<sub>2</sub>O || 5.25 || 7.25 | |||
|- | |||
| || 10.0 || 10.0 | |||
|} | |||
Thermal cycler | |||
* [45°C, 2 min.; 16°C, 5 min.] x25 | |||
* 60°C, 20 min. | |||
* 80°C, 20 min. | |||
---- | ---- |
Revision as of 17:46, 16 January 2013
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01/15/13
Golden Gate Assembly optimization
1, 2. Promega - hPCD+BL01+pSB1A3
Thermal cycler
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