Streptococcus Growth Curve Protocol

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1. Turn on the Bio safety level 2 hood, and spray down with ethanol

2. Spray gloves with ethanol and wipe inside of hood down

3. Steps 4-6 should be conducted in the hood

4. Set up 6 13mm tubs labeled S. Pneumonia 1-6 and add 3ml of Tryptic soy broth media to each

5. Remove the aliquots from the -20 degree freezer and let them thaw

6. Pipet 30ul of the aliquots into each tube and vortex

7. Leave the tubes in 34 degree, 5% CO2 incubator for 2 hours

8. Remove the first tube, vortex (in the hood) and place in the spectrometer

9. Note: make use to have set up the spectrometer by blanking it

10. Record the absorbance and return the tube to the incubator

11. Later remove the first and second tube from the incubator

12. Vortex (in the hood) and place in the spectrometer

13. Record the absorbance and return tubes to incubator

14. Repeat this with each successive tube

Modified from: Iannelli, F. & Pozzi, G. Mol Biotechnol (2004) 26: 81.