Spring 11:Directed Evolution/Week of 4-18-2011 Status
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Revision as of 00:53, 28 April 2011 by Huayu Ding (Talk | contribs)
Contents
Weekly Progress
Weekend/Monday
- Finalized parts list.
- Continued coding image analysis / segmentation code.
- Fabricated Microfluidics using resources from Kamm lab and Voldemann lab.
- PDMS poured & cured Monday afternoon
- Moved microscope setup to larger breadboard.
Monday Evening
- 2 Cultures of I13521 started from freezer stocks tubes 2&3. Aliquoted into 5mL of LB+Amp and left to grow overnight. ~12 hours of growth time should be sufficient for experimentation tomorrow.
- Idea: potentially growing up KAF95 and miniprepping it may prove useful if we can BioBrick out the fliC273 gene.
Tuesday
- Aligned microscope setup on the larger breadboard
- Tested different perfusion chambers:
- PDMS with microwell is too thick to be imaged with inverted microscope
- Stick on perfusion chambers are reusable
- Visualized RFP bacterial cells under BF and Fluoresence, pictures were taken for further analysis
Wednesday
- Begin building stepper motor driver circuitry with Polulu and Arduino
- Experimenting with serial communication in Python, wrapper for stage control
- Arduino C language for firmware in microcontroller, buffers and interpret serial input in bytes, and output electrical pulses
Thursday
- Stage control code tested: stepper motor can run at different speeds with specified number of steps through python
- Input as (X direction, X step, Y direction, Y step)
- X-Y axes motors move simultaneously given input to desired location
Friday
- Mutation plasmid from Liu's group acquired. Transformed and grew in spectinomycin, but antibiotic was too weak, redo over weekend.
- Finalizing and committed stage code.
- Meeting with another MIT group to discuss building the ablation laser.
Notes
Biology
- David Liu has agreed to give us MP-QUR, pickup will be TBD, but we can hopefully just go over to Harvard and get the plasmid.
- The Elowitz Repressilator can apparently be found on Add-Gene, will need to either find or figure out a good testing rig to put it in.
- Cory has I13521 and I13522, although both are in pSB1A3, which might prove problematic if the KAF95 fliC is also on a pBR322 derived plasmid. Current backup is J04450 which can be found in pSB1C3 and would allow for a co-transformant.