phuong-#4 - Group Members_Date LI Unique Day Room Lab Start...

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Group Members: _______________________________________________________________Date: ___________ LI ______________________ Unique __________ Day: __________ Room: _______ Lab Start Time ________ BIO206L Spring 2011 Exercise 4 Analysis To be completed as a group and turned in at the beginning of your next laboratory period. Include your “Data & Results”, sketches, acquired digital images, etc. as directed by your laboratory instructor. Show your work for all calculations and/or print your MS Excel data sheets. Be sure to include proper units where necessary. Adhere to University’s Honor Code and course policies. Balance between brevity and completeness. (1) Understanding the basics of the polymerase chain reaction. (a.) For one of the PCR samples prepared by your group, list the contents of each tube and describe the role of each of the components found in the PCR reactions. Master Mix 12.5ul in all 7 PCR samples (a, b, c, d, e, and f), 2.5ul into a-c, 10ul of H2O in A, 10ul of the Human Genome in samples d, e, f, and g, 10ul of P-15 into sample b, Plasmid DNA -colony into sample a. (b.) What was the final volume of the reaction mix in the tube? Sample a = total of 25ul Sample b = total of 25ul Sample c = total of 25ul Sample d = total of 25ul Sample e = total of 25ul Sample f = total of 25ul Sample g = total of 25ul (c.) What was the final concentration of the 2X Taq Buffer? The final concentration of the Taq Buffer is 1X. This is because the Taq makes up half of the total contents, thus the original concentration is divided by 2. (d.) What is the function of the Chelex beads during genomic DNA isolation? What is the effect of Chelex beads on the subsequent PCR reaction using the genomic DNA you isolated? The Chelex increases the binding ability of Mg+2 to decrease the DNAse activity. (2) The rate of polymerization of Taq polymerase is 1000 bp/min. Your supervisor has asked you to PCR amplify an interesting DNA sequence that is of normal GC content and 1.5 kb in length. (a.) What elongation/polymerization time would you use in your PCR program?
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phuong-#4 - Group Members_Date LI Unique Day Room Lab Start...

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