BME100 s2015:Group9 12pmL4: Difference between revisions
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<!-- Include a description of the thermal cycling program below. You can use text, a screen capture, a camera snapshot of the computer screen, or a digital drawing (e.g., using shapes and text boxes in Microsoft Powerpoint) --> | <!-- Include a description of the thermal cycling program below. You can use text, a screen capture, a camera snapshot of the computer screen, or a digital drawing (e.g., using shapes and text boxes in Microsoft Powerpoint) --> | ||
Heated Lid: 100°C | Heated Lid: 100°C <br> | ||
Initial Step:95°C for two minutes | Initial Step:95°C for two minutes <br> | ||
Number of Cycles: 35 | Number of Cycles: 35 <br> | ||
Denature at 95°C for 30 seconds, Anneal at 57°C for 30 seconds, Extend at 72°C for 30 seconds. | Denature at 95°C for 30 seconds, Anneal at 57°C for 30 seconds, Extend at 72°C for 30 seconds. <br> | ||
Final Step: 72°C for 2 minutes | Final Step: 72°C for 2 minutes <br> | ||
Final Hold: | Final Hold:4°C <br> | ||
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'''PCR - The Underlying Technology'''<br> | '''PCR - The Underlying Technology'''<br> | ||
<!-- 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. --> | ||
<br> | |||
Question 1: <br> | |||
Template DNA: The strand of DNA that will serve as the beginning structure in order to become a complete double helix in DNA synthesis. <br> | |||
Primers: A strand of nucleic acid that serves as a starting pint in PCR aslo reffered as base nucleotides. <br> | |||
Taq Polymerase: Its a thermostable DNA polymerase that is an enzyme in this process of replicating the complementary nucleotides.<br> | |||
Deoxyribosenucleotides(dNTP's): The monomer that is compromises into three parts which is one phosphate group, deoxyribose sugar and a nitrogenous base. <br> | |||
<br> | |||
Question 2: <br> | |||
Initial Step: <br> | |||
The current research of the PCR where they are are using a template DNA sample where there is one original sample of DNA from a hair follicle, blood, skin and etc to be the start of the process. <br> | |||
Denature: <br> | |||
After setting it up in a deoxyribose nucleoside triphosphate solution mix (dNTP) in which serve as nucleotide building blocks and are essential to maintaining life, the next step is heating it up the solution to 95°C in time for denaturing the double helix into two individual strands. <br> | |||
Anneal: <br> | |||
Right after stability is achieved, the temperature is then switched to 57°C to add strands of nucleic acid or primers where they anneal to complementary matches on the two DNA strands where they bracket themselves into prepared sequences of the template DNA. <br> | |||
Extension: <br> | |||
Next comes the step of raising the temperature to 72°C and then adding the enzyme Taq Polymerase to the primed sequences. <br> | |||
Final Step: <br> | |||
The enzymes then add the remaining nucleotides to make a complete second copy. In which we have a pair of double helix DNA which are exact copies of each other <br> | |||
Final Hold(Repetition if necessary): <br> | |||
Then this first cycle can be repeated over and over again until the required amount of copied DNA is reached. <br> | |||
<br> | |||
Question 3: <br> | |||
Base Annealing Combinations: <br> | |||
Adenine(A): Thymine(T)<br> | |||
Thymine(T): Adenine(A)<br> | |||
Cytosine(C): Guanine(G)<br> | |||
Guanine(G): Cytosine(C) <br> | |||
<br> | |||
Question 4: <br> | |||
Base-pairing occurs during the "Anneal" and "Extend" steps of thermal cycling in which these nucleotide bases help finish creating a double helix DNA. | |||
<br><br> | <br><br> | ||
[[Image:PCR lab.jpg]] | |||
"Polymerase Chain Reaction | Biochemistry." Encyclopedia Britannica Online. Encyclopedia Britannica, n.d. Web. 01 Apr. 2015. | |||
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Latest revision as of 13:15, 1 April 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
have the same forward primer and reverse primer
samples will be cross-contaminated
OpenPCR program Heated Lid: 100°C
Research and DevelopmentPCR - The Underlying Technology
"Polymerase Chain Reaction | Biochemistry." Encyclopedia Britannica Online. Encyclopedia Britannica, n.d. Web. 01 Apr. 2015. |