Protocol for the First Experiment 3/2/2012

Protocol

  1. Obtain bacterial culture with pre-transformed bacteria (with gene that synthesizes CdS)
  2. Prepare subculture of bacteria cells into 200 mL of LB broth, OD600 = .6 at 37C
  3. Prepare 6 samples: no bacteria control, no IPTG control, 3hr CdCl2, 2hr CdCl2, 1hr CdCl2 and a 3hr CdCl2 with no Na2S
  4. The below procedure is based off of the summer iGEM procedure
  5. Induce culture with .5 mM IPTG for 4 hours
  6. Take 10 mL of LB medium with kanamycin
  7. Add CdCl2 (approximately .00183g) until the final concentration is 1 mM
  8. Incubate the sample for 3 hours (check progress each hour)
  9. Add freshly prepared Na2S 9H20 (approximately .024018g) to final concentration 1 mM
  10. Incubate samples at room temperature with end-over-end rotation for 1.5 hours
  11. Wash final sample with water 3 times, place resulting sample under UV-vis and fluorescence spectrometry

Possible derivatives to experiment

  1. Set negative control of experiment
    • Repeat steps 1-9 without the initial bacteria
    • This procedure ensures that the bacteria is synthesizing CdS (quantum dot)
  2. Adjust the times of incubation
    • Adjust the the total times of incubation (suggested by steps 6 and 8)
    • Adjust by plus/minus 1 or 2 hours
    • Adjustment to see if incubation time affects the productivity in synthesis of CdS

Material Safety Data Sheet

Material Safety Data Sheet

Note: Protocol is based off experiment conducted by Mi et al.
Article source: Biosynthesis and characterization of CdS quantum dots in genetically engineered E. Coli from Journal of Biotechnology

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start/classes/principlesofdesign/igem/experiment_1_protocol.txt · Last modified: 2012/03/02 15:09 by ymakita
 
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