Team:ICT-Mumbai/Experiments

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Protocols

Bacillus subtilis transformation protocol

1. Streak a large patch of Bacillus subtilis cells onto an LB agar plate (2.5% LB and 1% agar)
2. Incubate at 37 deg C overnight
3. Scrape cell’s off the patch into 10 ml of freshly made SpC medium (for contents of SpC medium see below) till OD of tube becomes approximately 0.5
4. Incubate at 37 deg C and 220 rpm for 5 hours
5. Inoculate 200ul of this culture into 10 ml of freshly prepared SpII medium (for contents of SpII medium see below)
6. Incubate at 37 deg C and 170 rpm for 90 min
7. Pellet cells in 1.5 ml microcentrifuge tubes. Decent and store supernatant
8. Resuspend pellets in 200ul of supernatant
9. Add 15ul of plasmid (pBS1C backbone)
10. Incubate at 37 deg C
11. Plate on LB agar plates (2.5% LB and 1% agar) containing chloramphenicol (25 ug/ml)
12. Incubate overnight at 37 deg C to grow colonies

Why is the sky blue?

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Will we ever discover aliens?

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How much does the Earth weigh?

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How do airplanes stay up?

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