BIOC301 – Xmas Review Problem Set
Q1. Which of the following primers would allow for extension of the single-
stranded DNA molecule 5’-GTACTAACCGA-3’ ?
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’
Ina OVR-h 5’-ATGGTACTAACCGAGATGCCGCAAATAGATTAA-3’
Primer 1 (5’à3’): ____________________
Primer 2 (5’à3’): ____________________
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.
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
Q5. A 1:10 solution of 100.0mg/mL PMSF (MW 174.2) is of what molarity? Show
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)
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