Bio Lab Practical Exam

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66 Terms

1
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measuring absorbance: 1

labquest app, sensors, calibrate

2
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measuring absorbance: 2

90 sec warm up, blank cuvette in, finish calibration

3
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measuring absorbance: 3

sensors, data collection, time based mode

4
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measuring absorbance: 4

red box > select wavelength, change absorbance

5
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measuring absorbance: 5

cuvette in blank, play, 10 seconds > stop

6
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measuring absorbance: 6

analyze, statistics, record avg (mean) absorbance

7
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measuring absorbance: 7

cuvette 1 in cuvette 0 place, play, discard data

8
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calculating pH: 1

probe into channel 1, rinse w/ distilled water, wipe w kim wipe

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calculating pH: 2

submerge probe, wait 15 sec before data collection, play button > collect data for 10 sec

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calculating pH: 3

stop button, analyze, statistics, record avg pH

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calculating temperature: 1

temp probe into channel 1, stick probe/buffer into diff environments

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calculating temperature: 2

play, collect data 10 sec, stop, analyze, statics, record avg temp

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total solids

if solids mass at least 0.025 > after subtracting final - initial mass, proceed

14
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BOD: 1

DO probe into channel 1, rinse tip w/ distilled water, wipe w kim wipe

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BOD: 2

switch on probe to mg/L, submerge for 40 sec, collect data for 10 sec, stop

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BOD: 3

analyze, statistics, avg DO concentration, record under initial dissolved oxygen

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BOD: 4

still submerged, flip switch to %, press play, measure 10 sec, rec avg % saturation

18
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Nitrates: 1

clamp probe, insert nitrate ISE to clamp vertically, uncap nitrate high standard, place o support stand directly under nitrate ISE

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Nitrates: 2

lower probe clamp to submerge nitrate ISE in nitrate high standard (NOT touching bottom), soak for 30 min

20
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Nitrates: 3

ISE sensor into channel 1 of vernier interface, sensors, calibrate, select nitrate ISE

21
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Nitrates: 4

“calibrate now”, wait for live voltage stabilization, value 1 = 100, keep, swap nitrate high standard for rinsate beaker

22
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Nitrates: 5

rinse, swap rinsate beaker for nitrate low standard, lower clamp so nitrate ISE submerged into low standard

23
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Nitrates: 6

wait for live voltage stabilization, know value 2 = 1, keep, finish calibration, rinse ISE

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Nitrates: 7

ISE tip into Nalgene bottle, probe submerged 60 sec, collect data 10 sec, stop collection, analyze, statistics, rec avg nitrate concentration

25
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total phosphate: 1

calibrate spectrovis, connecct to labquest via USB, sensors, calibrate, spectrometer, 90 sec warmup

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total phosphate: 2

blank in cuvette holder of spectrovis, finish calibration, ok, red box, change wavelength, enter given absorbance, ok

27
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proteins

dependent on pH, temp, salt concentration of reaction mixture

28
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buffers

aqueous solution mix, function to resist changes in hydrogenn ion concentration

29
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stock solutions

concentrated solutions that last over a long period

30
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dilution factor

concentration of dilute solution reduced compared to concentration of stock solution

31
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serial dilution

stepwise dilution where stock solution for each dilution is dilute solution from previous dilution

32
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spectrophotometer

machine measures absorbance or transmittance of pigmented solutions

33
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standards (solution)

have known amount of material assayed, used to calculate amount in unknowns

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standard curve

determine concentration of unknown

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independent variable

x axis, not affected by other variables

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dependent variable

y axis, influenced/changes in response to independent variable

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discovery science

uses data or surveys of natural systems to discover patterns/correlations

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hypothesis-driven science

uses scientific method to develop knowledge

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descriptive statics

describe/summarize data

-mean, median, mode measure central tendency

-range/variance/standard deviation measure dispersion

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inferential statistics

make estimates/draw conclusions/inferences about population based on sample (T-Test)

41
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temp effects on WQI

-cold water = more dissolved gas/DO

-temp changes indicate source of thermal pollution

-increased temp increases photosynthesis

-organisms stressed, lowering resistance

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thermal pollution

caused by human activities

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runoff

increases overall temp of water

44
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shade

important to stream health, blocks too much direct sunlight

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pH effects of WQI

algal blooms, industrial processes, sulfide sediment oxidation, rainfall

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total solids effects on WQI

-soil erosion

-runoff

-plankton/animal and plant matter

-dissolved solids dehydrate organisms

-high total solids > cloudy water > less photosynthesis

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factors affeting DO concentrations

temp, stream flow, air pressure, aquatic plants, decaying organic matter, human activities

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BOD effects on WQI

-Oxygen replenished faster than used

-too much organic material to decompose = high BOD

-initial DO - final DO = BOD level

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Nitrate effects on WQI

-increased by fertilizers, decomposition, waste, runoff, etc.

-high levels cause eutrophication

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Total phosphate effects on WQI

-high levels associated w decreased DO/increased BOD

-added by humans

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Fecal coliform bacteria, WQI

-indicated other pathogens from feces

-caused by leaking sewer systems, polluted runoff, etc.

-can survive wo oxygen

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Turbidity, WQI

-increase w stream flow, soil erosion, runoff, etc.

-caused by bottom dwelling organisms, organisms, plankton

53
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Conserved DNA region

-nucleotide seq w little/no variation across species, remains constant throughout evolution

54
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Bacteria DNA barcoding

-agar as growth medium

-no buffer, prokaryotic

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DNA purification/isolation: 1

Heat sample to 94 degrees celcius for 5 min, purify DNA, centrifuge Lysate

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DNA purification/isolation: 2

DNA binds column, wash buffer clears everything else away (4x), nuclease free water added to column/centrifuged, DNA pulled through column/isolates

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PCR

-Denaturing: DNA in microcentrifuge for 1 min at 95 degrees celcius

-annealing

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Master mix

-DNA polymerase, dNTPs, buffer, forward/reverse primers, water

-reduced pipetting errors/time running reaction

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Gel electrophoresis: 1

-separate, identify, purify DNA frags

-DNA migrates to wards + charged electrode, smaller move faster through matrix

-Agarose/buffer cool/hardening jello substance

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gel electrophoresis: 2

-blue gel dye added, EtBr added for frag visualization

-gel run at 175 volts, turn off when migrated 5 cm

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gel analysis: presence of ladder

gel made properly, electrophoresed in correct direction, stained by etbr

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gel analysis: no bands including ladder

etbr not added

63
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gel analysis: ladder visible but bands not

part of taq polymerase degraded maybe

64
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gel analysis: no sample band

one or both primers not added to master mix, DNA extraction recovered little/no genomic DNA

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gel analysis: no band in bacteria

did not properly inoculate bacteria culture

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gel analysis: multiple PCR bands

non-specific primer binding, sample contamination, primer added to multiple sample. master mix