User:Karmella Haynes/Notebook/BioBrick cloning/2009/11/18

{| width="800"
 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] Project name
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


 * colspan="2"|
 * colspan="2"|

11/18/09

 * &#x2713; Cultures: KAH77(2), 78(3), 79(3), 83(2), 84(2), 85(2)
 * &#x2713; Assemblies: KAH86, 87, 88, 89, 90, 91, 92
 * &#x2713; Oligo annealing: NLS myc stop (KAH011), K-stop (KAH012), BglII adapters

Assemblies
 * 1) KAH86: HSVtkTATA-K (KAH48)/(S/P)/~3200 &#x2713; + (1) 2xYFP-NLSstop2-polyA-2xHS4 (KAH81)/(X/P)/2640
 * 2) KAH87: (2) huPcCD/(E/S)/186+3200 + (8) mCh-VP64 (KAH74)/(E/X)/4061
 * 3) KAH88: (3) mPcCD/(E/S)/186+3200 + "
 * 4) KAH89: (4) flyPcCD/(E/S)/186+3200 + "
 * 5) KAH90: (5) fshPcCD/(E/S)/186+3200 + "
 * 6) KAH91: (6) hCBX8/(E/S)/1170 + "
 * 7) KAH92: (7) flyPc/(E/S)/1173 + "

> Digests (Fermentas FD) --> Gel purify 1, 6, 7, 8 --> Zymo clean & conc. 2-5 (for dephos.)

> Measure conc.'s

> Dephosphorylation (Roche)

> Ligations (*Note: use insert + old backbone in calculations)

Oligo annealing
 * 1) NLS myc stop (KAH011)
 * 2) K-stop (KAH012)
 * 3) Adapter BglII/EcoRI
 * 4) Adapter PstI/BglII


 * }