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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 Procedures—make an instruction page on how to pour plates Safety Precautions wear gogles Tutorials 13 14 15 17 18 media prep. lb agar and lb broth introduction to sterile technique streaking for isolated colonies streaking for broth culture Vocabulary • • • • • • • • • • • 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 Mass 1/vol 1 = mass 2/ volume 2 Lab 4E tips 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 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
