IGEM:MIT/2007/Notebook/2007-7-16: Difference between revisions
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# CPX - At FedEx Facility in Boston | # CPX - At FedEx Facility in Boston | ||
# FhuA - ? | # FhuA - ? | ||
==Sequencing of LC#1&2 from 3A== | |||
*Miniprepped LC's of those colonies that grew only on Kan plates and not on Kan/Amp plates | |||
*Sent in 4 samples, one forward and one reverse for each LC | |||
*used 4µl of DNA since each was approximately 30ng/µl | |||
Revision as of 08:29, 16 July 2007
F2620 Expression Protocol
- Create Liquid Cultures in LB Media
- Dilute Back 1:500 in LB Media
- Grow up for an hour in warm room roller
- Split culture in two
- Add AHL to one culture to a final concentration of 1E-6M
- Grow up for 4 hours in warm room roller
- Spin Down, Decant, and Resuspend in PBS
- Measure absorbance at 600nm and red fluorescence level
See characterization data for BBa_F2620.
Today's Agenda
- Redo Overgrown Liquid Cultures
- Send for Sequencing
- Look for AHL Induced Expression
- Cut CPX and Backbone for ligation
- Start designing high copy backbone
- PCR off relevant portion of Mer operon from pto40
- Organize fridge
- Design and order EC_20 Insert
- Email Isadora + Tom Knight for Meeting Times--Toan--Done
- Anneal FHUA Insert oligos
Status of Expected Parts
- IDT Primers + Inserts - Arriving Today--ARRIVED
- CPX - At FedEx Facility in Boston
- FhuA - ?
Sequencing of LC#1&2 from 3A
- Miniprepped LC's of those colonies that grew only on Kan plates and not on Kan/Amp plates
- Sent in 4 samples, one forward and one reverse for each LC
- used 4µl of DNA since each was approximately 30ng/µl
