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Transcript of Topic 1 Tools of the Biology Laboratory Exercise I. The spectrophotometer. Exercise II. Liquid...
![Page 1: Topic 1 Tools of the Biology Laboratory Exercise I. The spectrophotometer. Exercise II. Liquid Measurements. Exercise III. Sedimentation and Centrifugation.](https://reader035.fdocuments.us/reader035/viewer/2022062500/5697bff91a28abf838cbf94e/html5/thumbnails/1.jpg)
Topic 1Tools of the Biology Laboratory
Exercise I. The spectrophotometer.
Exercise II. Liquid Measurements.
Exercise III. Sedimentation and Centrifugation.
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Bio 185 Labs: goal
When:
Self-paced:
Lab-Report:
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Bio 185 Labs: set-up.
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Exercise I.The Spectrophotometer
Experiment 2: Absorption Spectra of Dyes.
Experiment 1: Concentration of an unknown Dye.
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Exercise I. The Spectrophotometer
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Spectrophotometry:
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Spectrophotometry:
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Exercise I. The Spectrophotometer
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Exercise I. The Spectrophotometer
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Exercise I. The Spectrophotometer
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Exercise I. The Spectrophotometer
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Exercise I. The Spectrophotometer
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Spectrophotometry:You need to “Blanc” the spectrophotometer for each solvent and change of wavelength.
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Exercise I: Experiment 2: Absorbance spectrum:
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Absorbance spectrum:
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Absorbance spectrum:
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Exercise I. The Spectrophotometer: Absorption spectrum.
1. Change wavelength. 2. Blanc. 3. Measure.
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Exercise I. The Spectrophotometer: Absorption spectrum.
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What wavelength to determine the concentration of a solution?
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Exercise I. The Spectrophotometer: Concentration of an unknown dye.
Standards of known protein concentration
Optical density of solutions with unknown concentration.
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LAB REPORT FORMAT: (1 report per group!)
LAB TITLE: Concentration of a dye.Experiment 1.
Goal: one sentence.Method: Relevant.Results: (based on class data). A (mean and SEM) is significantly different
(P<0.05) from B (mean and SEM).Conclusion: A is different from B.Explanation.
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Standard Curve Yellow Dye
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1 1.5 2 2.5 3 3.5 4
Concentration (drops)
Op
tica
l Den
sity
at
420
nm
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Figure 1. Standard curve of yellow dye at 420 nm. We measured the optical density at 420 nm of four concentrations of yellow dye (1, 2, 3, and 4 drops per sample) using a Spectronix 21 spectrophotometer. The OD of Standard 1 ((0.217 ± 0.0070) was significantly smaller (P<0.05, Student’s t-test) than that of Standard 2 (0.410 ± 0.0057), the OD of Standard 2 was significantly smaller (P<0.05) than that of Standard 3 (0.619 ± 0.0045) and the OD of Standard 3 was significantly smaller (P<0.05) than that of Standard 4 (0.806 ± 0.0087). We calculated the standard curve (linear regression y = 0.1977x + 0.0186, R2 = 0.9996).We measured the optical density at 420 nm of an unknown concentration of yellow dye (OD equals 0.699 ± 0.0054) and used the standard curve to determine its concentration (3.44 drops per sample).
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Exercise II.Liquid Measurements
Accuracy of (1) 100 ml Graduated cylinder, (2) 10 ml serological pipette, and (3) 1.0 ml micropipette.
Percent error when device is used at maximal and 1/10th maximal nominal volume.
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Exercise II. Liquid Measurements
Measure by weight (mass).
Read mass in g.Tare or zero the balance.Add volume of water.Measure.
What mass (g) do you expect?
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Exercise II. Liquid Measurements: 50 ml cylinder.
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Exercise II. Liquid Measurements: 10 ml pipette.
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Exercise II. Liquid Measurements
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Exercise II. Liquid Measurements: 1.0 ml micropipettor.
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Exercise II. Liquid Measurements
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Exercise II. Liquid Measurements
100 ml 10 ml 10 ml 1 ml 1 ml 0.1 mlcylinder serological pipet micropipetor
0
4
8
Accuracy of Volumetric Devices.
Volumetric Devices
Pe
rce
nt
Err
or
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Exercise III. Sedimentation and Centrifugation.
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Exercise III. Sedimentation and Centrifugation.
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Exercise III. Sedimentation and Centrifugation.
Sedimentation at 1 g Sedimentation in 50 ml conical tube at slow centrifugation
Sedimentation in microfuge tubes at high speed centrifugation
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Exercise III. Sedimentation and Centrifugation.
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Exercise III. Sedimentation and Centrifugation.
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Exercise III. Sedimentation and Centrifugation.
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Exercise II. Liquid Measurements
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Exercise II. Liquid Measurements
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Exercise II. Liquid Measurements
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Exercise III. Sedimentation and Centrifugation.
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Exercise III. Sedimentation and Centrifugation.
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Microscope:
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Exercise III. Sedimentation and Centrifugation.
Sedimentation at 1 g Sedimentation in 50 ml conical tube at slow centrifugation
Sedimentation in microfuge tubes at high speed centrifugation