Beruflich Dokumente
Kultur Dokumente
REVIEW
Thursday June 14 during class
LAB MATH: Conversions
Reminders:
■ 1 L = 1000 mL
■ 1 mL = 1000 microliters
■ Assume 1 g = 1 mL
■ : and / are used interchangeably for describing dilution ratios
LAB MATH: Weight/Volume%
mass of solute (g)
w/v% concentration = x 100
volume of solution (mL)
EXAMPLE: How many grams of salt would you need to make 1L of a 0.9% NaCl
solution?
1000 𝑚𝐿
1𝐿 𝑥 = 1000 mL
1𝐿
𝑥 𝑔𝑟𝑎𝑚𝑠
𝑥 100 = .9%
1000 𝑚𝐿
𝑥 𝑔𝑟𝑎𝑚𝑠
= .009
1000 𝑚𝐿
𝑥 = 9 g NaCl
LAB MATH: Molar Concentration
moles (mol) grams
Molarity (M)= Molecular weight (MW)=
liter (L) 1 mole
1 1 1 1 1 1 .5 3 1 1 2
𝑥 𝑥 𝑥 = 𝑥 𝑥 𝑥 𝑥 𝑥 =
10 10 10 10 10,000 10 5 30 100 10 20
Serial dilution! 1 1 1 1 1 1 1
𝑥 𝑥 𝑥 𝑥 𝑥 =
10 10 10 100 10 10 10,000,000
EXAMPLE: How would you make 240mL of a 1:3 dilution of dye?
1 x
=
3 240 𝑚𝐿
𝐼𝐶 1
𝑥 = 10,000
𝐹𝐶 1/10,000
C1V1 = C2V2
x = 2% dextrose solution
LAB MATH: Diluting Solutions (Unspecified Volumes)
EXAMPLE: How do you dilute a solution that has an initial concentration of
solute of 10% down to 2%?
Assume 1 L of initial
10% x 1 L = 2% x X L solution. OR:
𝐼𝐶 10%
AMOUNT of solute in 1 L 𝐷𝐹 = = =5
10% x 1 L = 2% x 5 L of 10% is equal to 𝐹𝐶 2%
AMOUNT in 5L of 2%.
Reminders:
■ Never “redip” from your original
plate!
■ Reflame between each quadrant
■ Wait until loop cools/stops glowing
red
■ Use Bunsen burner/aseptic
technique
Good!
Contamination (not a pure culture)
Week 2
Pipetting
Reminders:
■ Volumes should only be set from
X/10 – X microliters
– Example: P100 should only
be set from 10-100
microliters
■ Keep pipette upright when
there’s liquid inside
■ Check for bubbles in pipette tip
Week 2
Bacterial Growth
𝑡𝑖𝑚𝑒
Generation time =
𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑔𝑒𝑛𝑒𝑟𝑎𝑡𝑖𝑜𝑛𝑠
𝑡2 −𝑡1 𝑥 log(2)
Generation time = 𝑂𝐷𝑡2
log( )
𝑂𝐷𝑡1
MasterMix:
Elongation TAQ Polymerase
72° C dNTPs
dNTPs added 16S rRNA primers
Sterile PCR water
Cathode (-)
Week 3
Gel Electrophoresis
■ Check PCR product
■ Smaller DNA: move faster
■ Larger DNA: moves slower
Anode (+)
Week 3
Bioinformatics
■ BLAST
– Identify organism by nucleotide sequence
– Use FASTA format
■ Gideon
– Identify organism by biochemical test results
– Input both positive/negative results!
■ Phylogeny.fr
– Know how to read relationships from tree
Week 4
MRSA
■ Methicillin Resistant Staphylococcus aureus
■ 25-30% population colonized, but not necessarily infected
■ Transmitted in hospital + community
■ Produce beta-lactamase resistance to penicillin class drugs
■ Infection starts off as small pimple-like bump + often develops pus
drainage; causes boils, abscesses, and furuncles
Week 4
ELISA (Enzyme-Linked Immunosorbent Assay)
Week 5
Antibiotics