Microbial Ecology and Evolution Lab Wiki

General microbiology protocols

Cloning and gene manipulation

Commonly Used Plasmids

Plasmid Purification

Digest and Ligation

Creating Competent E. coli Cells

Transformation

Gibson Assembly

Phyllosphere protocols

Creating Sterile Agar Plates

Sterilization and Germination Protocol for ''Arabidopsis thaliana'' Seeds in Gnotobiotic Experiments

Germination Protocol for ''Arabidopsis thaliana'' Seeds in Non-Sterile Experiments

Growth Stage Phenotype Definitions

Growth Conditions for ''Arabidopsis thaliana''

Measuring Light with HOBO Data Loggers

Inoculation of ''Arabidopsis thaliana'' with Microbes

Removal and DNA Extraction of Phyllosphere Microbes

ARISA

Measuring ''A. thaliana'' Phenotype using FIJI

Streptococcus pneumoniae protocols

Dual Layer Assays

Streptococcus DNA Extraction

Streptococcus Growth Curve Protocol

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. https://doi.org/10.1385/MB:26:1:81

Streptococcus suis protocols

Streptococcus suis Transformation

Measuring Absorbance in Streptococcus

Streptococcus DNA Extraction

Streptococcus Competence Induction

Peptide Synthesis

Peptide Cleavage

Mass Spectrometery

Plate Reader Assay and Growth Curve

Measuring Competence : Fixation and Flow Cytometry

Remote Molecular Biology

Effect of Laboratory Protocols on Student Learning

Cambridge protocols

Storage buffer

transformation of R5(2)-mCh-FL-BST and

expression

lysis and immobilization

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