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Midterm

BIO314H5 Study Guide - Midterm Guide: Differential Centrifugation, Centrifugal Force, Particle Velocity


Department
Biology
Course Code
BIO314H5
Professor
Steven Chatfield
Study Guide
Midterm

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Lecture 1
SI system; prefixes
Making solutions
Easy way:
Weight-in-weight (w/w)
Weight-in-volume (w/v)
Volume-in-volume (v/v)
Not always useful:
If one wants EQUAL amounts of molecules, weight is almost never a good way to achieve it
Making solutions – molarity
Describe how you would make 500 mls of a 1M NaCl solution. MW NaCl = 58.44g (MW =
molecular weight; synonym is FW = formula weight)
1 Mole of NaCl weighs 58.44 g
1 M = 58.44 g/L
To make 500 mL
500mL/1000mL x 58.44g = 29.22g
Therefore add 29.22g to 400mL and after it dissolves bring the volume to 500mL
Diluting solutions

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Making dilutions
Have a 1.0 Molar stock solution
Want 10 ml of a 10 mM solution
c1V1=c2V2 or cHVH=CWVW
1.0M x VH = 0.01M X 10mL
VH = 0.01M x 10mL/ 1.0M = 0.1 mL = 100μL
Therefore add 100μL of 1M solution to 9.9 mL of water
Making stock solutions
What solvents will my compound dissolve in?
oHow soluble?
Which solvent is appropriate for my experiment?
oE.g. will it kill my cells in minute concentrations?...
How can I make my life easier?
oE.g. 1000X stocks ?
oE.g. Enough for my entire PhD?
Storage and perishability?
When using a centrifuge make sure:
To consult the appropriate manual and/or a colleague officially entrusted with demonstrating
its use before using any centrifuge for the first time.
To have the rotor speed and force matching the maximum allowable for the given rotor type,
tube type, and task at hand.
To have the rotor positioned in right place, tightly screwed on the centrifuge shaft, and lid
placed on the top.
To balance the opposing loads (1g makes a big difference at 20,000G)
To stay next to the centrifuge until the operating speed is reached.
Rotor types
Centrifuge disasters can result in:

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Serious bodily injuries (and death)
Contamination (biohazard, radioactivity, chemicals)
High lab costs
oUltracentrifuge: ~100-200K$
oTitanium rotor: ~15-30K$
oPreparative centrifuge (floor): ~40-60K$
oPreparative centrifuge (bench): ~30-50K$
oTechnician per hour: ~150-300$
Loss of material, time
You will be “that guy” or “we had this student…”
Rotor corrosion and stress
Physics 101
Angular velocity is the speed at which something rotates. The SI unit of angular velocity is radians
per second (rads-1), but other units such as revolutions [=rotations] per minute (RPM) are in
common use. Angular velocity is usually represented by .ω
Centrifuge rotors
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