实验动物模型

Response to food assayby Meng-Qiu Dong 1/1/2000Wild type worms adjust egg-laying behavior in response to food. They lay eggs when they are fed and stop laying eggs when they are starved. If starved w ...

Mutagenesis of C. elegansMaterials:ENG plates 0.5% Peptone 0.1% Yeast Extract 51 mM NaCl 2% agaradd after autoclaving: 5 ug cholesterol 1 mM CaCl2 1 mM MgSO4 1 mM K-phosphate (pH 6) 1 M NaOH make up 5 ...

EMS MutagenesisWARNING:EMS is poweful mutagen and a suspected carcinogen. Wear gloves and work in fume hood. Use disposable plasticware and inactivate mutagen before disposal as outlined below.1. Grow ...

F2 Mutant Screen - non "clonal"by Michael Koelle adapted from Erik Jorgenson4/6/941. Mutagenize mixed population worms as per normal. Best to do this first thing in the morning in order to get several ...

Fermentor worm chowPrep Have autoclaved: 250 ml straight neck flask 100 mmfunnel Fermentor vessel with all openings covered top loosened on its side in tub with wedges after a/c sit upright to seal a ...

Cleaning worms by floating on sucroseHave cold 70% sucrose in water. Wash worms off plates put on ice.Typically we use 2 - 10 10-cm plates of crowded or starved wormsFor this many worms use a 15ml ora ...

Materials:disposable scapel #15 (round tip) Feather brand subbed slides 0.1 % poly-L-lysine (MW300000) 0.2 % gelatin 0.01 % chrome alum (dip slides in solution and dry overnight) Whatmann paper for ea ...

Freezing Worms by Michael Koelle 4/6/94 I. The preferred method 1. Wash worms off of 1 large plate or 3 small plates in 3 mls S Basal into a sterile 15 ml disposable centrifuge tube. Worms should be h ...

Gene Knockout with Conventional MutagensLeon AveryENG (enriched nematode growth medium): 1. in a 6 L flask add: 3800 ml DH2O 20 g. bactopeptone 4 g yeast extract 12 g. NaCl 4 ml 5mg/ml cholesterol in ...

Growing worms in liquidFirst make worm food -- grow liters of bacteria Inoculate each of several liters (4 is a good number) of LB in 2 liter flasks with 1 ml of an overnight culture of AMA1004. Grow ...

Integrating extrachromasomal arrays into the C. elegans chromosomes Why and how to do it by Michael Koelle 4/6/94 What is the benefit of integrating an extrachromosomal array? Extrachromosomal arrays ...

Lethal phase determination1. Pick a bunch of heterozygote L4 hermaphrodites to a new plate.2. The next day there will be eggs on the plate; pick exactly 20 of these to each of a few plates. Make sure ...

Integrating extrachromasomal arrays into the C. elegans chromosomesWhy and how to do itby Michael Koelle12/20/94What is the benefit of integrating an extrachromosomal array? Extrachromosomal arrays su ...

Microinjecting worms by Michael Koelle8/23/94You will have a couple frustrating sessions when you first attempt this technique but everyone seems to master injection after a few days and it works very ...

Generating males by heat shockby Michael Koelle1. Set up ~6 plates with 5 L4 hermaphordites each.2. Heat shock 4-6 hours at 30°. 8-9 hours works but gives few progeny.3. Return to 20°. Should get a fe ...

LIQUID CULTURE OF WORMS WITH FERMENTORMimi H. Zhou Hsiuchin Yang Bill Walthall Biology Department Georgia State University Atlanta GA 30302 The yield of C. elegans and their food (E.coli) are relative ...

Liquid culture of wormsBy Michael Koelle and Tory Herman adapted from Mir Hengartner4/6/94Media1) superbroth (5 X 2 1/2 liters autoclaved in 6 liter flasks) 5 X 30 g Bactotryptone 5 X 60 g yeast extra ...

Single Worm PCRReagents:Lysis Buffer: 50 mM KCl10 mM Tris pH 8.32.5 mM MgCl20.45% NP-40 (IGEPAL)0.45% Tween-200.01% Gelatin Proteinase K: 20 mg/ml 10X PCR Buffer: 100 mM Tris 500 mM KCl 15 mM MgCl2 pH ...

snip-SNP mapping with CB4856 polymorphismsMaterials:Worm Lysis Buffer PCR reagents PCR primers flanking a SNP 2X restriction mix with the appropriate enzyme Procedure:Generate recombinant lines for th ...

Synchronizing Worm CulturesSynchronization is based on the fact newly hatched larvae will live but not develop if deprived of a food source. An asynchronous population of embryos is isolated and then ...