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Biochemistry
DAY 5: Identification of Unknown RFP Introduction Every fluorescent protein has unique biophysical properties that can be characterized spectroscopically using different complimentary techniques. Even fluorescent proteins of the same apparent 'color' (as judged by your eye) can have dramatically different absorption and fluorescent properties. For each fluorescent protein, these properties are known and well characterized, and can be found in many review articles and in primary scientific literature. The goal for today's lab is to spectroscopically characterize your unknown RFP to determine the identity by comparing to known values for mCherry, mPlum, and TagRFP. Specifically, you will need to design a set of experiments that will allow you determine which RFP you have based on absorption and fluorescent properties of your unknown. Assigned Reading • Guide to Choosing Fluorescent Proteins (Canvas) You will need to find articles that give biophysical data about mCherry, mPlum, and TagRFP to help you set up a experimental protocol to determine the identity of your unknown RFP. There are many search engines that you can use, but Google (www.google.com) and PubMed (www.ncbi.nlm.nih.gov/pubmed/) are good places to start (Hint: try using keywords such as: RFP, review, fluorescent protein, TagRFP, mCherry, mPlum, choosing fluorescent protein). You should be able to find multiple references, which you will need for your formal report.

DAY 5: Identification of Unknown RFP Introduction Every fluorescent protein has unique biophysical properties that can be characterized spectroscopically using different complimentary techniques. Even fluorescent proteins of the same apparent 'color' (as judged by your eye) can have dramatically different absorption and fluorescent properties. For each fluorescent protein, these properties are known and well characterized, and can be found in many review articles and in primary scientific literature. The goal for today's lab is to spectroscopically characterize your unknown RFP to determine the identity by comparing to known values for mCherry, mPlum, and TagRFP. Specifically, you will need to design a set of experiments that will allow you determine which RFP you have based on absorption and fluorescent properties of your unknown. Assigned Reading • Guide to Choosing Fluorescent Proteins (Canvas) You will need to find articles that give biophysical data about mCherry, mPlum, and TagRFP to help you set up a experimental protocol to determine the identity of your unknown RFP. There are many search engines that you can use, but Google (www.google.com) and PubMed (www.ncbi.nlm.nih.gov/pubmed/) are good places to start (Hint: try using keywords such as: RFP, review, fluorescent protein, TagRFP, mCherry, mPlum, choosing fluorescent protein). You should be able to find multiple references, which you will need for your formal report.

Biology 2e
Biology 2e
2nd Edition
ISBN:9781947172517
Author:Matthew Douglas, Jung Choi, Mary Ann Clark
Publisher:Matthew Douglas, Jung Choi, Mary Ann Clark
Chapter4: Cell Structure
Section: Chapter Questions
Problem 25CTQ: In your everyday life, you have probably noticed that certain instruments are ideal for certain...
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The electronic excitation method in fluorescence and absorption spectroscopy is the same. What are some of the variations between using a fluorimeter to perform an excitation scan and an absorption scan? Do you think the two spectra should be similar? Describe.

 

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DAY 5: Identification of Unknown RFP Introduction Every fluorescent protein has unique biophysical properties that can be characterized spectroscopically using different complimentary techniques. Even fluorescent proteins of the same apparent 'color' (as judged by your eye) can have dramatically different absorption and fluorescent properties. For each fluorescent protein, these properties are known and well characterized, and can be found in many review articles and in primary scientific literature. The goal for today's lab is to spectroscopically characterize your unknown RFP to determine the identity by comparing to known values for mCherry, mPlum, and TagRFP. Specifically, you will need to design a set of experiments that will allow you determine which RFP you have based on absorption and fluorescent properties of your unknown. Assigned Reading • Guide to Choosing Fluorescent Proteins (Canvas) You will need to find articles that give biophysical data about mCherry, mPlum, and TagRFP to help you set up a experimental protocol to determine the identity of your unknown RFP. There are many search engines that you can use, but Google (www.google.com) and PubMed (www.ncbi.nlm.nih.gov/pubmed/) are good places to start (Hint: try using keywords such as: RFP, review, fluorescent protein, TagRFP, mCherry, mPlum, choosing fluorescent protein). You should be able to find multiple references, which you will need for your formal report.
Transcribed Image Text:DAY 5: Identification of Unknown RFP Introduction Every fluorescent protein has unique biophysical properties that can be characterized spectroscopically using different complimentary techniques. Even fluorescent proteins of the same apparent 'color' (as judged by your eye) can have dramatically different absorption and fluorescent properties. For each fluorescent protein, these properties are known and well characterized, and can be found in many review articles and in primary scientific literature. The goal for today's lab is to spectroscopically characterize your unknown RFP to determine the identity by comparing to known values for mCherry, mPlum, and TagRFP. Specifically, you will need to design a set of experiments that will allow you determine which RFP you have based on absorption and fluorescent properties of your unknown. Assigned Reading • Guide to Choosing Fluorescent Proteins (Canvas) You will need to find articles that give biophysical data about mCherry, mPlum, and TagRFP to help you set up a experimental protocol to determine the identity of your unknown RFP. There are many search engines that you can use, but Google (www.google.com) and PubMed (www.ncbi.nlm.nih.gov/pubmed/) are good places to start (Hint: try using keywords such as: RFP, review, fluorescent protein, TagRFP, mCherry, mPlum, choosing fluorescent protein). You should be able to find multiple references, which you will need for your formal report.
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