User:Karmella Haynes/Notebook/BioBrick cloning/2010/01/25

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 * style="background-color: #F2F2F2" align="center"|  |Main project page


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01/25/10

 * &#x2713; Colony PCR screen: KAH 94, 95/ JDS48, 49; HS4 primers
 * &#x2713; Assembly: KAH140, 141/ V0201 (re-do, d.p. vector)
 * &#x2713; Cultures: KAH94/JDS48 #1 & KAH94/JDS49 #18 (2 each)

Colony PCR screen > Use BB_HS4 f1 & BB_HS4 r1 primers > Controls: (+) = KAH94 (0.5μL); (-) = JDS48 (0.5μL)

--> BioRad 96-well:
 * 95°C/ 3 min.
 * [95°C/ 30 sec., 57°C/ 30 sec., 72°C/ 30 sec.] x30
 * 72°C/ 3 min.
 * 4°C/ ∞

--> Check minipreps of KAH94/JDS48 #1 & KAH94/JDS49 #18 tomorrow with BglII and PstI digests

Assemblies
 * 1) KAH140/V0201: KAH140/(N/S)/2945 + V0201/(N/S, d.p.)/5157
 * 2) KAH141/V0201: KAH141/(N/S)/2939 + "

> Dephosphorylation (Roche) --> d.p. treat V0201(N/S)

> Ligations


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