User:Karmella Haynes/Notebook/BioBrick cloning/2010/08/07

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08/07/10

 * &#x2713; Assemblies: KAH160-170 into KAH015 (lentivirus vector)

Assemblies
 * 1) KAH160/KAH015: (1) KAH160/(E/S)/1147 + (12) KAH015/(E/X)/8402
 * 2) KAH161/KAH015: (2) KAH161/(E/S)/1147 + "
 * 3) KAH162/KAH015: (3) KAH162/(E/S)/1147 + "
 * 4) KAH163/KAH015: (4) KAH165/(E/S)/2131 + "
 * 5) KAH164/KAH015: (5) KAH164/(E/S)/2134 + "
 * 6) KAH165/KAH015: (6) KAH165/(E/S)/991 + "
 * 7) KAH166/KAH015: (7) KAH166/(E/S)/991 + "
 * 8) KAH167/KAH015: (8) KAH167/(E/S)/991 + "
 * 9) KAH168/KAH015: (9) KAH168/(E/S)/1955 + "
 * 10) KAH169/KAH015: (10) KAH169/(E/S)/1958 + "
 * 11) KAH170/KAH015: (11) KAH170/(E/S)/955 + "

> Digests (Fermentas FD)

> Measure conc.'s

> Dephosphorylation (Roche) --> KAH015(E/X)

> Ligations

--> R.T./~10 min.; Add to 50 μL T-DH5α cells

8/08/10 --> Many colonies on the ligation plates, none on the negative control. Looks great! Pick one colony from each plate; confirm via digest.


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