User:Karmella Haynes/Notebook/BioBrick cloning/2013/02/23: Difference between revisions
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| pSB1A3 || 2000 || 85.5 || 12.2 || 27.8 | | pSB1A3 || 2000 || 85.5 || 12.2 || 27.8 | ||
|- | |- | ||
| H2B || 378 || 40.4 || 4.9 || | | H2B || 378 || 40.4 || 4.9 || 35.1 | ||
|- | |- | ||
| LOV+his || 450 || 18.6 || 12. | | LOV+his || 450 || 18.6 || 12.6 || 27.4 | ||
|} | |} | ||
Line 88: | Line 88: | ||
# H2B + LOV+his + pSB1A3 | # H2B + LOV+his + pSB1A3 | ||
# pSB1A3 | # pSB1A3 | ||
# H2B + LOV+his + pSB1A3, 2xDNA | |||
# pSB1A3, 2xDNA | |||
{| {{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 || Master mix (x3) | | Reagent || 1 || 2 || Master mix (x3) || 3 || 4 || Master mix (x3) | ||
|- | |- | ||
| gg2 pSB1A3 || 1.0 || 1.0 || --- | | gg2 pSB1A3 || 1.0 || 1.0 || --- || 2.0 || 2.0 || --- | ||
|- | |- | ||
| gg3 LOV || 1.0 || H2O || --- | | gg3 LOV || 1.0 || H2O || --- || 2.0 || H2O || --- | ||
|- | |- | ||
| gg4 H2B+his || 1.0 || H2O || --- | | gg4 H2B+his || 1.0 || H2O || --- || 2.0 || H2O || --- | ||
|- | |- | ||
| 10x PRO ligase buffer || 1.0 || 1.0 || 3.0 | | 10x PRO ligase buffer || 1.0 || 1.0 || 3.0 || 1.0 || 1.0 || 3.0 | ||
|- | |- | ||
| NEB T4 lgase || 0.25 || 0.25 || 0.75 | | NEB T4 lgase || 0.25 || 0.25 || 0.75 || 0.25 || 0.25 || 0.75 | ||
|- | |- | ||
| BsmBI || 0.5 || 0.5 || 1.5 | | BsmBI || 0.5 || 0.5 || 1.5 || 0.5 || 0.5 || 1.5 | ||
|- | |- | ||
| dH<sub>2</sub>O || 5.25 || 5.25 || 15.75 | | dH<sub>2</sub>O || 5.25 || 5.25 || 15.75 || 2.25 || 2.25 || 6.75 | ||
|- | |- | ||
| || 10.0 || 10.0 | | || 10.0 || 10.0 || || 10.0 || 10.0 || | ||
|} | |} | ||
* Aliquot 7.0 mix to each tube; add DNA/ H2O | * Aliquot 7.0 mix to each tube; add DNA/ H2O |
Revision as of 12:23, 25 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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