Practical Study Guide.pdf

Practical Study Guide.pdf - EXERCISE Learning Objectives...

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EXERCISE Learning Objectives Ex 1: Introduction to Biology, Microscopy, and Pipettes Know how to use PPE appropriately. 1. Identify proper and improper PPE 2. ALSO WEAR YOUR PPE to the practical Know (both compound and dissecting) microscope parts and their function. 1. Identify the parts of a microscope 2. Know the function 3. Similar to the last practical Know how to safely operate both a compound and dissecting microscope 1. Know the steps of how to properly use a microscope (page 16, 20, and 22) 2. Be able to demonstrate this Know how to safely and properly aspirate using a microliter pipette 1. Page 4 describe proper technique 2. Rewatch the video on GauchoSpace 3. Be able to demonstrate this Know the volume limitation of different size microliter pipettes. (page 4) 1. Know the volume ranges for each pipette 2. Know the how to read the volume on the dials of a pipette 3. Know how to set the correct pipette if given a volume Ex 2: PCR Amplification of an Alu Insert & Analysis Using Agarose Gel Electrophoresis Know components of a PCR reaction & describe how PCR amplifies DNA. 1. What is required in a PCR tube? (Page 29) a. Understand the role of each component 2. What are the stages/steps of PCR? (Page 29) a. What is occurring during each step? Know how to create PCR master-mixes. 1. Find on page 32, step 9 Know how to set up and perform DNA (agarose gel) electrophoresis 1. Rewatch the video on GauchoSpace on how to step up a box for gel electrophoresis (be able to demonstrate this) 2. Know how to calculate how much agar powder you need for a specific percent gel (Page 39, question 4) Know how to visualize PCR products following gel electrophoresis.
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1. Normally, you cannot see DNA with the naked eye. How do we see it in gel electrophoresis? Know how to interpret gel electrophoresis results. 1. Given a gel, be able to calculate Rf. 2. Understand what different PCR bands mean for having or not having the Alu insert Ex 3: Biochemical Analyses of a Crude Homogenate: (i) Determining Total Protein Concentration of a Homogenate & (ii) Quantifying Tyrosinase Enzymatic Activity Present Within the Homogenate •Know the principles behind spectrophotometry and how to operate a spectrophotometer. (page 51) 1. Understand the difference between absorbance and transmission. 2. How does a spectrophotometer read absorbance? •Know how to generate a standard curve and how it is used to determine the protein concentration of an unknown solution. (Page 53-57) 1. What is the point of a standard curve?
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