其他模型

Preparation of Copy Standards for PCR GenotypingPCR screens must be designed to detect transgene DNA at the single copy level. To demonstrate this level of sensitivity non-transgenic tail DNA is spike ...

Preparation of Mouse Tail DNA for Dot Blots or PCRThese procedures were originally devised in Richard Palmiter's lab for use with tail dots (DNA spotted onto a nitrocellulose filter and probed for a t ...

Tail Preps: DNA Isolation From Mouse Tails Without PhenolNOTE: THIS IS FINE FOR SOUTHERNS BUT NOT FOR SCREENING BY PCR. (from Ruixia 7/99 from protocol by Stef Oehen 7/94). 1. Put 1 cm tail in 1.5 ml ...

DETECTION OF ß-GALACTOSIDASEThe E. coli lacZ gene encoding ß-galactosidase (ß-gal) is the classical histochemical reporter gene (Beckwith 1980). It can be detected using a variety of substrates all o ...

RNA-isolation (TRIZOL method)Isolation of RNA from whole worms using TRIZOL reagent (Gibco-BRL).1) Wash worms from a 60 mm plate with 1 ml DEPC-treated water.2) Spin down worms at 4000xg in microfuge ...

Nematode Genomic DNA IsolationMaterials:Rich Agarose Plates: 50 mM NaCl 0.75 % BactoPeptone 5 ug/ml cholesteroundefined 1 mM CaClundefined 1 mM MgSOundefined 25 mM K-PO4 (pH 6.0~undefined 1.5 % agarose (not agar) * add after auto ...

Cleaning Worm Stocksby Michael Koelle4/6/94There are two kinds of contaminants on worm plates:1. Fungi: these contaminants can come from the plates or bacteria so it is best to leave plates out after ...

Overview Using this procedure it is possible to follow the development of pollen mother cells through to mature pollen. Sterile males were made using irradiation mutagenesis of Landsberg erecta seed ...

Simplified Arabidopsis Transformation Protocol(Brief version for those who are familiar with the method)Steve Clough and Andrew Bent University of Illinois at Urbana-Champaign. Our present protocol (C ...

Culturing Wormsby Michael Koelle1. Bacterial strain for feeding worms: OP50 a uracil auxotroph. Streak OP50 out on a 9 cm NGM agar plate grow overnight at 37°. Can then parafilm the plate and keep it ...

Procedure1. Immerse Arabidopsis seeds in 10% Household Bleach for 20 min. 2. Rinse the seeds twice with 500 ml of sterile ddH2O and allow them to dry overnight in a laminar flow hood. 3. Push the res ...

Green lab protocol for vacuum infiltration transformation of Arabidopsis Step by step picture demonstration This protocol is adapted from protocols by Nicole Bechtold (Bechtold et al ...

Development of techniques for primary culture of C. elegans embryonic neurons Laird Bloom MIT from Ph.D. thesis Massachusetts Institute of Technology 1993 Introduction One of the major limitations of ...

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 ...