LAB FINAL REVIEW
Your lab final will assess your understanding of the lab experiments that you carried out this
semester. You will have approximately 10 –15 written questions and one “hands on portion
“where you will demonstrate a technique.
For each lab make sure you can answer the following questions:
● What is the purpose of this lab?
● What are the key biological concepts someone would need to understand this lab?
● Can I describe the general steps of this lab?
● What important techniques or assays did we use in this lab?How do they work and why
did we use it?
General Knowledge to Know:
1) Know how to use a micropipette.
2) Know how to dilute a stock solution to make a working solution (For instance by using C1V1
= C2V2 )
3)Know how to use a spectrophotometer and how you would go about making and using a
standard curve.
4) Know what molecule or entity you measured in each experiment and what you used to
measure it
5) Know the purpose of each reagent used in the lab (dye, buer, enzyme, etc.)
6) Know how to use, and why you use, a molecular weight marker (aka ‘ladder’).
7) Understand the need for negative and positive controls and be able to identify the controls in
your experiments
9) Know why a p value is important
10) Know how to use proper units (For instance, the symbol for ‘microliter’ is ‘μl’ )
Specific Topics:
1) The analysis of protein and glucose/glycogen in organ homogenates. 2) Membrane transport
using red blood cells as a model system.
3) Succinate dehydrogenase activity in organ homogeneates.
4) Assaying photosynthesis and respiration.
5) The transformation of E. coli, the positive selection of plasmids, the detection of DNA
sequences, and the detection of gene expression.
, Lab 1: Micropipette and Dilution proficiency
a. Purpose: To learn how to transfer volumes in the microliter range (using a
micropipette) and knowing how to make a dilution.
b. Key concepts:
i. Remember that 1 milliliter (mL) = 1000 ul
ii. Correct pipettes to use include the following: (Note: Bold ones are
important)
1. P2: 0.5-2 ul
2. P10: 0.5-10 ul
3. P20: 2-20 ul
4. P100: 10ul-100ul
5. P200: 20-200 ul
6. P1000: 100-1000ul
c. General steps:
i. Add tip
ii. Push to first stop
iii. Place into solution + Release Plunger
iv. Place tip into M.C. Tube + push plunger to second stop
d. Techniques: Honestly just know how to hold a micropipette (Thumb on top knob,
finger should cover the number window).
i. An example would be 950 ul. You would use the P1000 ul and there three
places are as follows: Thousands, Hundreds, Tens.
ii. DILUTIONS: Use C1V1 = C2V2
1. Given a concentration and volume: ex) Dilute a 10x solution of
Red dye to a 500μL, 1X Red Dye solution. How much Red Dye
and water will be in your diluted sample? (10x)(?)=(1x)(500ul)
2. To dilute given a ratio: Make a 1:5 dilution of homogenate with
water in a total volume of 250ul. (250 ul/5 = 50 ul) Since 50ul is
the homogenate, 200 ul is H2O.
Lab 2: Analysis of Protein in Bos taurus Homogenates
● Purpose: Measure and compare glycogen and protein from tissue homogenates from Bos
taurus.
● Key concepts: QUANTITATIVE AND QUALITATIVE
○ Biological molecule: Protein which have 20 amino acids joined by covalent
peptide linkages to form chains
○ Spectrophotometer: instrument designed to detect amount of radiant energy
absorbed by the molecules of a solution
Your lab final will assess your understanding of the lab experiments that you carried out this
semester. You will have approximately 10 –15 written questions and one “hands on portion
“where you will demonstrate a technique.
For each lab make sure you can answer the following questions:
● What is the purpose of this lab?
● What are the key biological concepts someone would need to understand this lab?
● Can I describe the general steps of this lab?
● What important techniques or assays did we use in this lab?How do they work and why
did we use it?
General Knowledge to Know:
1) Know how to use a micropipette.
2) Know how to dilute a stock solution to make a working solution (For instance by using C1V1
= C2V2 )
3)Know how to use a spectrophotometer and how you would go about making and using a
standard curve.
4) Know what molecule or entity you measured in each experiment and what you used to
measure it
5) Know the purpose of each reagent used in the lab (dye, buer, enzyme, etc.)
6) Know how to use, and why you use, a molecular weight marker (aka ‘ladder’).
7) Understand the need for negative and positive controls and be able to identify the controls in
your experiments
9) Know why a p value is important
10) Know how to use proper units (For instance, the symbol for ‘microliter’ is ‘μl’ )
Specific Topics:
1) The analysis of protein and glucose/glycogen in organ homogenates. 2) Membrane transport
using red blood cells as a model system.
3) Succinate dehydrogenase activity in organ homogeneates.
4) Assaying photosynthesis and respiration.
5) The transformation of E. coli, the positive selection of plasmids, the detection of DNA
sequences, and the detection of gene expression.
, Lab 1: Micropipette and Dilution proficiency
a. Purpose: To learn how to transfer volumes in the microliter range (using a
micropipette) and knowing how to make a dilution.
b. Key concepts:
i. Remember that 1 milliliter (mL) = 1000 ul
ii. Correct pipettes to use include the following: (Note: Bold ones are
important)
1. P2: 0.5-2 ul
2. P10: 0.5-10 ul
3. P20: 2-20 ul
4. P100: 10ul-100ul
5. P200: 20-200 ul
6. P1000: 100-1000ul
c. General steps:
i. Add tip
ii. Push to first stop
iii. Place into solution + Release Plunger
iv. Place tip into M.C. Tube + push plunger to second stop
d. Techniques: Honestly just know how to hold a micropipette (Thumb on top knob,
finger should cover the number window).
i. An example would be 950 ul. You would use the P1000 ul and there three
places are as follows: Thousands, Hundreds, Tens.
ii. DILUTIONS: Use C1V1 = C2V2
1. Given a concentration and volume: ex) Dilute a 10x solution of
Red dye to a 500μL, 1X Red Dye solution. How much Red Dye
and water will be in your diluted sample? (10x)(?)=(1x)(500ul)
2. To dilute given a ratio: Make a 1:5 dilution of homogenate with
water in a total volume of 250ul. (250 ul/5 = 50 ul) Since 50ul is
the homogenate, 200 ul is H2O.
Lab 2: Analysis of Protein in Bos taurus Homogenates
● Purpose: Measure and compare glycogen and protein from tissue homogenates from Bos
taurus.
● Key concepts: QUANTITATIVE AND QUALITATIVE
○ Biological molecule: Protein which have 20 amino acids joined by covalent
peptide linkages to form chains
○ Spectrophotometer: instrument designed to detect amount of radiant energy
absorbed by the molecules of a solution
