User:Karmella Haynes/Notebook/BioBrick cloning/2009/12/22

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12/22/09

 * &#x2713; Cultures: KAH120-125 in MV2 (2 each); KAH135 (2)
 * &#x2713; Digests/ dephos.: Vectors for Strept apt into JDS42 & 43
 * &#x2713; Kinase treatment + oligo annealing: Strept apt

Assemblies
 * 1) Strep apt/JDS42: Strept apt/(NotI dsoligo)/50 + (1) JDS42/(NotI)/7468
 * 2) Strep apt/JDS43: Strept apt/(NotI dsoligo)/50 + (2) JDS43/(NotI)/7478

> Digests (Fermentas FD)

> Measure conc.'s

> Dephosphorylation (Roche)

Kinase treatment & oligo annealing
 * 1) Joe's: Strep apt S1 F NotI + Strep apt S1 R NotI

> Kinase treatment

--> 37°C/ 1.5 hours --> Add 4 uL 500 mM NaCl --> 100°C (water bath)/ 5 min.; Cool to R.T. overnight in water bath


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