BME100 s2015:Group5 9amL4: Difference between revisions
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'''Q4: During which two steps of thermal cycling does base-pairing occur? Explain your answers. | '''Q4: During which two steps of thermal cycling does base-pairing occur? Explain your answers. | ||
Base-pairing occurs during the extended and "final step" parts of PCR. Taq polymerase is optimized during the extended phase and can attach to the primers of the template DNA strands, and during the final step the proper base pairs are gathered and attached to their correct partners. | |||
<!-- Add a write-up, essay-style, organized into paragraphs with descriptive headers, based on the questions and answers from the Research and Development exercise. BONUS points: Use a program like Powerpoint, Word, Illustrator, Microsoft Paint, etc. to illustrate how primers bind to the cancer DNA template, and how Taq polymerases amplify the DNA. Screen-captures from the PCR video/ tutorial might be useful. Be sure to '''credit the sources''' if you borrow images. You are not allowed to use images from current or past BME 100 students' reports on OpenWetWare. --> | <!-- Add a write-up, essay-style, organized into paragraphs with descriptive headers, based on the questions and answers from the Research and Development exercise. BONUS points: Use a program like Powerpoint, Word, Illustrator, Microsoft Paint, etc. to illustrate how primers bind to the cancer DNA template, and how Taq polymerases amplify the DNA. Screen-captures from the PCR video/ tutorial might be useful. Be sure to '''credit the sources''' if you borrow images. You are not allowed to use images from current or past BME 100 students' reports on OpenWetWare. --> |
Revision as of 20:07, 31 March 2015
BME 100 Spring 2015 | Home People Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3 Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6 Course Logistics For Instructors Photos Wiki Editing Help | ||||||||||||||||||||||||||||||||
OUR TEAMLAB 4 WRITE-UPProtocolMaterials
Research and DevelopmentPCR - The Underlying Technology Q1: What is the function of each component of a PCR reaction Template DNA: This is the sequence of DNA that is intended for amplification. It will undergo extreme heat in order for it to denature. Primers: A primer is a single strand of DNA that serves as the starting point for DNA synthesis that binds to specific DNA sequences. Taq Polymerase: This is a thermostable enzyme that recombines nucleotides following separation by heat. It creates a new, complimentary strand of DNA through adding new nucleotides to the old ones. Deoxyribonucleotides (dNTP's): There are the building blocks of DNA: adenine (A), thymine (T), cytosine (C) and guanine (G). These are used by the Taq Polymerase to synthesize the complementary strand of DNA. Q2: What happens to the components (listed above) during each step of thermal cycling? Initial step (95°C for 2 minutes): The temperature of the reaction is increased to 95°C and held for two minutes in preparation for separation. Denature at 95°C for 30 seconds: The double stranded DNA in the reaction is broken down into single strands of DNA for amplification. Anneal at 57°C for 30 seconds: The temperature of the reaction is lowered to around 5°C below the melting point of the primer (57°C) so that the primers will bind to specific points on the template DNA that indicate areas that will be amplified. Extend at 72°C for 30 seconds: The temperature of the reaction is increased to the ideal temperature for polymerase activity (72°C) so that the Taq polymerase will bind to the primers that are already on the template DNA and will begin to synthesize the complimentary strand. Final step (72°C for 3 minutes): These three minutes are used to make sure that each strand has been fully extended. Final hold (4°C): This is the temperature to deactivate the Taq polymerase and end the process. Q3: DNA is made up of four types of molecules called nucleotides, designated as A, T, C and G. Base-pairing, driven by hydrogen bonding, allows base pairs to stick together. Which base anneals to each base listed below?;;;
Q4: During which two steps of thermal cycling does base-pairing occur? Explain your answers. Base-pairing occurs during the extended and "final step" parts of PCR. Taq polymerase is optimized during the extended phase and can attach to the primers of the template DNA strands, and during the final step the proper base pairs are gathered and attached to their correct partners.
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