Identifying bacterial species is important for the following reasons: Disease diagnosis: Identifying pathogens can help with diagnosing diseases. Treatment: Identifying pathogens can help with treating infections. Food safety: Identifying contaminants can help ensure food safety. Safety hazards: Identifying bacteria can help with safety hazards. Does sealing Petri plate influence bacterial growth? http://dx.doi.org/10.13140/RG.2.2.20537.44641 Check out my RG profile for various bacterial identification reports https://www.researchgate.net/profile/Chinnamani-Prasannakumar https://www.youtube.com/watch?v=T4DRexF4Mms

1. Mother plate First quadrant streak
colony characterization
First glycerol stock Second quadrant streak Second glycerol stock Third glycerol stock
Gram’s staining Oxidase test Catalase test Broth
Broth characterization
Motility check at 18th hour Antibiotic sensitivity test
Spore staining IMViC assay TSI test
Carbohydrate fermentation test:
Minimum of 3 carbohydrates Growth curve
Remember to
document the
results of
each and
every
experiment

2. Characterization of bacteria
Macro- & micro-
morphological analysis
Chemotaxonomy Molecular taxonomy
 Colony morphology
 Cell shape and size
 Mobility
 Total cellular protein fingerprinting
 16S rRNA gene sequencing
 Restriction digestion
 Extracellular polysaccharide profiling
 Cell wall composition
 KoH assay
 Oxidase
 Catalase
 IMViC
 TSI
 Carbohydrate fermentation
 Urease test
 Nitrate reduction test
 Antibiotic sensitivity assay

4. Pour plate Spread plate
Same soil sample: @10-5 dilution

6. Mother plate First quadrant streak
colony characterization
First glycerol stock Second quadrant streak Second glycerol stock Third glycerol stock
Gram’s staining Oxidase test Catalase test Broth
Broth characterization
Motility check at 18th hour Antibiotic sensitivity test
Spore staining IMViC assay TSI test
Carbohydrate fermentation test:
Minimum of 3 carbohydrates Growth curve
Remember to
document the
results of
each and
every
experiment

7. NCT45 = Tax. Score = 0.692
Tax. Score = sum of all score / no. of assay

17. Capsule and negative staining

19.  Butt should be 40% length of the tube.
 Slant should be in equal length to butt (up to 80% length of the tube).
 Way of inoculation: stab & streak
Triple sugar iron test
Glucose (0.1%)
Sucrose (1%)
Lactose (1%)

21. Observed within 15
seconds

28. 68 KDa
1
1
2
3
4
0
0
0
1
2
3
4
NCT27 = BSA004
1
1
1
2
3
4
5
6
7
8 NCT70 = BSA118

29. Include student name at the bottom of the gel
Student who did the isolation, not the gel.

31. NCT45 = Tax. Score = 0.692
Tax. Score = sum of all score / no. of assay

32. Step 1 Step 2 Step 3
Visualization of the
experiment before
entering the lab
Execution of the
experiment
Cleaning before
leaving the lab