enriched media
not selective or differential
difference between streak,
spread, and pour plate and
when to use
30-300
Gram (+) vs Gram (-)
Gram-variable
Cell Wall-less
cell vs colonial morphology
bacterial cell arrangement:
Strepto vs staphylo
selective vs differential
media
bacterial identification vs
detection
molecular approach vs
conventional
methods of sterilization
catalase test
bacterial cell count;
dilution 0.1 mL plating;
cells per mL vs total cell
count
antibiotic action vs
antibiotic resistance
in general, allows almost all bacteria to grow;
eg. L-agar/broth
valid count of CFUs
G-variable: accurate G staining cannot be obtained, may be misleading
CW-less Mycoplasma
chains & clusters
difference wrt purpose
identification: primarily uses 60S (bacteria) or 80s (if yeast/euk);
additional markers include gyraseB, rpoB;
16S rRNA and 18S rRNA
detection: conventional: biochemical tests
molecular approach: PCR. primer should target a gene that is unique/specific to the
species that you want to detect.
to confirm whether this was indeed the gene that was amplified: sequence
conventional: morphology; biochemical tests to characterize unknown bacteria
how to make media, glassware, etc free of contaminating organisms
autolave most commonly used temp and pressure
filter sterilization size of filter pore
UV targets nucleic acids (DNA, RNA)
when plating: flame the wire loop / needle
+ H2O2 (hydrogen peroxide) H2O + O2
if bacterium possesses catalase: bubble formation due to release of oxygen
distinguishes Staphylococcus (+) and Streptococcus (-)