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EXAM 2: MCB 2610 Exam 2 with Complete Solutions Cultivation independent methods CORRECT ANS-DNA from unculturable bacteria can be amplified and sequenced by PCR Sequences can be used to produce fluorescent probes that will bind to complementary DNA (Fluorescent In Situ Hybridization FISH) F...

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  • September 3, 2024
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  • 2024/2025
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  • exam 2 mcb 2610 exam 2 with complete solutions
  • cultivation independent methods correct ans dna

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EXAM 2: MCB 2610 Exam 2 with Complete Solutions
Cultivation independent methods CORRECT ANS-DNA from unculturable bacteria can be

amplified and sequenced by PCR


Sequences can be used to produce fluorescent probes that will bind to complementary DNA

(Fluorescent In Situ Hybridization FISH)




FISH CORRECT ANS-Cells fixed in place


Fluorescently labeled probes consisting of 165 rRNA gene sequence sent to attach to RNA


View in Epifluorescence microscope




Metagenomics CORRECT ANS-DNA is isolated from an environmental sample and

sequenced


Metagenomic information must still be confirmed in cultured organisms




Microbial consortia CORRECT ANS-Microbes that can't grow on their own




Direct counts CORRECT ANS-Load known volume into gridded slide


Count under a light microscope

, EXAM 2: MCB 2610 Exam 2 with Complete Solutions
Flow Cytometry CORRECT ANS-Microbial suspension forced through small orifice with a

laser beam


Movement of microbe through orifice impacts electric current that flows through orifice


Instances of disruption of current are counted


Specific antibodies can be used to determine size and internal complexity




Spread plate technique CORRECT ANS-used to get a living (viable) cell count


Small amount of sample is pipetted onto a solid medium then spread around that medium


Measured in CFU colony forming units




Pour Plate CORRECT ANS-Original sample is diluted multiple times and the most dilute

ones are poured into a plate with liquid agar


Isolated cells grow into colonies on the surface and inside of the medium


Measured in CFU colony forming units




Calculating CFU/mL CORRECT ANS-Average cell number / (dilution X volume plated)

, EXAM 2: MCB 2610 Exam 2 with Complete Solutions
What if cells are already very diluted? CORRECT ANS-A filter apparatus can concentrate

cells


Sample is filtered through a membrane filter then the contents that don't go through are

transferred to a medium and incubated




Turbidity CORRECT ANS-Spetrophotometer sends light through a liquid culture


If the tube is cloudy, light wont reach the other side because it will hit all the bacteria in the

tube


This can give a rough estimate of cell density in the tube


Higher absorbance = higher population in culture




Indirect measurements of cell mass CORRECT ANS-Dry weight - time consuming and not

very sensitive


Quantity of particular cell constituent - DNA protein ATP




Growth CORRECT ANS-Population growth vs growth of actual cells

, EXAM 2: MCB 2610 Exam 2 with Complete Solutions
Growth Curve CORRECT ANS-Observed when microorganisms are cultivated in batch

culture (closed culture)


Usually plotted as a logarithm of cell number vs. time


Has 4 distinct fases




Lag phase CORRECT ANS-Cell synthesizing new compounds


ex. to replenish spent materials, adapt to new conditions or medium




Exponential phase CORRECT ANS-Rate of growth and division is constant and maximal


Population is most uniform




Stationary phase CORRECT ANS-Closed system


Population growth eventually ceases


Reasons - nutrient limitation, limited oxygen availability, toxic waste accumulation - can make

environment too acidic




Stationary phase and starvation response CORRECT ANS-Activate survival strategies:


Morphological changes - endospore formation, decrease in size

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