User:Karmella Haynes/Notebook/BioBrick cloning/2011/02/22

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02/22/11

 * &#x2713; Gaussia luciferase: check minipreps with NotI digestion
 * &#x2713; Biobrick PCR: G luc

Gaussia luc minipreps > Check with NotI digest

BioBrick PCR --> Gaussia luciferase from Addgene22522 template
 * 1) KAH016: (1) Gluc/(X/P PCR)/558 + V0120/(X/P)/~3200 &#x2713;

> PCR --> Primers: BB_Gluc f/ BB_Gluc r

--> BioRad DNA Engine, block A
 * 95°C/ 3 min.
 * [95°C/ 30 sec; 55°C/ 30 sec; 72 °C/ 1 min.] x30
 * 72 °C/ 3 min.
 * 4 °C/ ∞

> Zymo clean PCR; elute w/ 25 μL dH2O

> Digests (Fermentas FD)

> Measure conc.'s

??? -- Nanodrop computer isn't working. Just use max volume insert for 10.0 μL reaction.

> Ligations

--> R.T./ ~15 min.; Add to 30 μL DH5α turbo


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