BIOC301 – Xmas Review Problem Set.pdf

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University of British Columbia
BIOC 301
Jason Read

BIOC301 – Xmas Review Problem Set Q1. Which of the following primers would allow for extension of the single- stranded DNA molecule 5’-GTACTAACCGA-3’ ? a) 5’-CATGAT b) 3’-CATGAT c) 5’-AGCCAA d) 5’-TCGGTT e) None Q2. Which of the following primers would allow for extension of the single- stranded DNA molecule 5’ –GCGGCCGC- 3’ ? a) 5’ – GCGG b) 3’- GCGG c) 5’ – GGCG d) 3’ – GGCG e) More than one choice Q3. The overshare-hydrolase(OVR-h) enzyme is inactivated in people who LUV fb & twitter. You postulate that this inactive (ina) form is different from the normal, wild-type (wt) form at the gene level and thus design an experiment to check this. Using the gene information below write out the primers you will use in your PCR reactions, 8mers should be sufficient here. wt OVR-h 5’-ATGGTACTAACCGAGCGGCCGCAAATAGATTAA-3’ 3’-TACCATGATTGGCTCGCCGGCGTTTATCTAATT-5’ Ina OVR-h 5’-ATGGTACTAACCGAGATGCCGCAAATAGATTAA-3’ 3’-TACCATGATTGGCTCTACGGCGTTTATCTAATT-5’ Primer 1 (5’à3’): ____________________ Primer 2 (5’à3’): ____________________ Knowing that the restriction endonuclease NotI cleaves after the first GC of the palindromic sequence 5’-GCGGCCGC, describe an experiment which would allow you to differentiate between inactive and active forms STARTING FROM PURIFIED DNA SAMPLES.     1   Q4. Your instructor has assigned you to clean up after a lousy TA who did not save PCR programs. You are to program the thermocycler so the BIOC420 class can proceed with their experiments this afternoon. In his notebook, you find some chicken-scratch and decipher the following: 72 / 1min 58 / .5min 94 / .5min 94 / 7min Repeat previous 3 steps 40x You suspect this information is out of order and re-input correctly into the thermocycler: Q5. A 1:10 solution of 100.0mg/mL PMSF (MW 174.2) is of what molarity? Show unit analysis. M Q6. You are volunteering as an undergraduate in Scott Covey’s lab. Because you successfully gone through the BIOC301 lab, he is confident you can make all of the solutions for his experiments. How would you make the following solutions? Note: you only have access to a weight-based scale; you CANNOT weigh out in mole units! a) 1L of 100mM Tris-HCL (121.14 g/mol) b) 200mL of .5M MgAc (110239 g/mol) c) 100μL of 1x Protease Inhibitor Cocktail (Stock from Roche Scientific is 25x) d) 416μl .25mM NADH (30mM stock solution)   2   Q7. Fischer Scientific is having a promotion and is selling EDTA (disodium salt, MW 416.2) at $10.00 per millimole. You tell your instructor and he asks you how much it will cost him for 500 lysis procedures. If you use 100ul of 0.5M pH8 for each lysate generated, how much will it cost to generate 500 lysates? $ Q8. You are to set
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