Team:Glendale CC AZ/Protocols/GrowthCurve

Glendale Community College Arizona

Protocols

Growth Curve Assay

NaCl Growth Curve Assay

Survival Growth Assay

Alkaline Lysis Plasmid Miniprep

Restriction Digest

DNA Isolation

Bioinformatics

Ligation

Transformation

E.coli Bacterial Growth Curve Assay Protocol

Materials

• 1 brain-heart infusion (BHI) broth culture of 5h E.coli
• 1 tube with 5.0 ml BHI broth
• Spectrophotometric tubes
• Plastic disposable pipettes
• Spectrophotometer at 590 nm 37°C water bath

Procedures

1. Add approximately 6 drops of the BHI into a spectrophotometer tube; this will be the blank tube.
2. Add about 6 drops of BHI into a spectrophotometer tube and and another 6 drops of the E.coli broth into the same tube; this will be experimental tube.
3. Read the blank tube on the spectrophotometer and blank the system at 590 nm.
4. Read the experimental tube at 590 nm and record the data.
5. Incubate the experimental tube for 20 minutes at 37°C.
6. Once the incubation is complete, read the experimental tube on the spectrophotometer and record the data.
7. Repeat steps 5-6  as many times as desired to get a good growth curve.
8. Graph the data accordingly.