User:Karmella Haynes/Notebook/BioBrick cloning/2010/07/14

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07/14/10

 * &#x2713; Cultures: pJ10's and pAct5C plasmid transformations (2 mL ~5 hr. Turbo cultures)
 * &#x2713; Assemblies: re-try cloning from yesterday (may have incorrectly cut V0120)

Minipreps > Check pJ10's with BamHI/KpnI digests > Check pAct5C with BioBrick enzymes:
 * 1) EcoRI/ XbaI
 * 2) NotI/ XbaI
 * 3) SpeI/ XbaI
 * 4) PsI/ XbaI
 * 5) XbaI
 * 6) 0 enzyme

--> Conclusions: pAct5C contains EcoRI and PstI, but not sure where since there is no sequence data. Will cut out renilla luc w/ SpeI/ NotI and replace with mini BB cloning site (NotI-SpeI). Order primers tomorrow.

Assemblies > See 07/13/2010

> Digests (Fermentas FD) --> Re-digest V0120

> Measure conc.'s

> Ligations

--> R.T./ 10 min.; Add 40 μL DH5α Turbo


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