Calendar:ImperialWetLabNotebook2009iGEM/2009-9-2

--Dineka Khurmi 08:57, 2 September 2009 (EDT)

WEDSDAY JOB LIST

• WaaL - Gel extraction - Run on gel with vector AK3 - Ligation

• Harvard BB - Gel extraction - Run on gel with GFP - Ligation

• CRP1 (vector) - Gel extraction - Run on gel with GFP - Ligation

• Miniprep cellulase - Digestion - Run on a gel

• Midiprep RFP I13507 (small culture)

• Dirty ligation RBS + AK3 - Transformation

• Pick culture colonies - grow 5ml

• Make agar for Amp/Kan plates and LB media (send to autoclave)

David Roche 05:55, 2 September 2009 (EDT)
AM : Ran a gel with the WaaL pcr products that were digested overnight. 20ul against the ladder
Updated wiki and BioBrick registry

• James Field 16:17, 2 September 2009 (EDT):

AM:

• Checked transformed plates from yesterday - only OtsA and Ots B had colonies (1 or 2 each). Positive control worked indicating that the ligation was

responsible for poor results.

• Did a mini Prep on the Edinburgh cellulases:

K118022

K118023

K118028

• Went through and made corrections to SLIC primers for Construct 9.

PM:

• Did Restriction digest on the Edinburgh cellulases and ran on gel. This indictated that the miniprep had been successful.

• Plated OtsA and OtsB colonies from the Ak3+insert ligation plates (see AM for more details). Also kicked off a starter culture for a mini prep for these colonies which will be conducted tomorrow.

• I transformed cells with the RBS + AK3 construct and plated out - will see results tomorrow following overnight incubation.

• Innoculated media (200ml LB) for overnight incubation for tomorrow's midi on:

RBS-RFP-TT (BBa_I13507)

K118022

K118023

K118028