Transcript Measuring small and very small volumes

Cell Cultures
Lab Manual 4e, 4f & 4g --Demo
timeline
Monday—Lecture
Tuesday—procedures quiz for all,
Perform Demo
Prep.
Tuesday
 Lb broth base and LB agar base
 Several 250ml jars
 Bring in extra hood
 10% Bleach
Lab Service
 Plates out
Write-up for 4e, 4f and 4g
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Procedures—make an instruction
page on how to pour plates
Safety Precautions
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wear gogles
Tutorials
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media prep.
lb agar and lb broth
introduction to sterile technique
streaking for isolated colonies
streaking for broth culture
Vocabulary
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Agar – solid media used for growing bacteria, fungi, plant, or other cells
Media preparation – the process of combining and sterilizing ingredients (salts, sugars,
growth factors, pH indicators, etc.) of a particular medium
Autoclave – an instrument that creates high temperature and high pressure to sterilize
equipment and media
Enhancer – a section of DNA that increases the expression of a gene
Intron – the region on a gene that is transcribed into an mRNA molecule but not expressed in
a protein
Exon – the region of a gene that directly codes for a protein; it is the region of the gene that
is expressed
Transcription factors – molecules that work to either turn on or off the transcription
eukaryotic genes
Histones – nuclear proteins that bind to chromosomal DNA and condense it into highly
packed coils
Nonpathogenic – not known to cause disease
Bacteriophages – viruses that infect bacteria
Gene therapy – the process of treating a disease or disorder by replacing a dysfunctional
gene with a functional one
equations
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Mass 1/vol 1 = mass 2/ volume 2
Lab 4E tips
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Clean and label glassware!
Media is hygroscopic
Slowly stir in small amounts
LB broth has: tryptone, yeast extract and NaCl
LB Broth 20g/1L—do the conversion for the amount
you are told to make
Use LB Broth base and add agar 1.5% to make LB
agar (Example: 20g/L broth and 15g/1L)
We will pour the plates next class
Keep lid loose
Bottle size needs to be twice the volume of recipe
Lab 4F tips
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Sterilize with 10% bleach spray
Re-heat LB agar in a microwave
pour plates in sterile environment
when LB agar has cooled, so you can
touch the glass
Leave plates in the hood
Plates will be used for lab 4g
Lab 4G tips
Bacteria waste goes into
sterilizer tray, so it can be
sterilized
 Clam method
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