实验动物模型

RNA In Situ Hybridization of Dissected Gonads with Digoxygenin-Labeled Probes- From R. Francis �- Revised by Min-Ho Lee 2003 -In contrast with other methods using gonads affixed to slide ...

Whole-mount in situ hybridization for the detection of RNA in C. elegans embryosGeraldine Seydoux and Andrew Fire Adapted from Seydoux G. and Fire A. (1995). Whole-mount in situ hybridization for the ...

Freeze-Crack P granule Staining Protocol1. Prepare humidity chamber: line large square plate with humid paper towels and place 4 5ml plastic pipets across chamber to support slides.Place aluminum bloc ...

PAR-1 Staining Procedure Freeze Fracture to permeabilize embryos Fixation 15' in -20 degree 100% MeOH1 x 10' wash in PBSStaining 1 x 60' block in goat serum1 x 60' primary antibody (rabbit-anti-PAR-1 ...

Sos ProtocolAntisera staining/ImmunohistochemistryFixationAMPA assayC. elegans cosmid vectors___________________________________________________________________________________Sos (volatile repellent ...

Phalloidin Staining worms by Michael Koelle modified from Beth Bucher and Andrew Chisholm 4/6/94 Phalloidin binds to filamentous actin. This stain allows visualization of mainly of muscles but some ot ...

Staining C. elegans for ß-galactosidase activity using X-gal by Michael Koelle adapted from Laird Bloom Jeff Way & Michael Basson 4/6/94 1. Grow up a small plate of worms. Some protocols call for usin ...

Protease Inhibitors from Alberts' Lab Protocols (Kathy Miller/Chris Field)Protease inhibitors are rapidly inactivated in aqueous solutions and should be added to solutions immediately before use. Cock ...

Stockkeeping1. MechanicsMost stocks can be successfully cultured by periodic mass transfer of adults to fresh food. Bottles or vials are tapped on the pounding pad to shake flies away from the plug t ...

I. MEDIUMA. S2 S2C Sundefined S2R+ S3 l(2)mbn Kc(167) & DL2 cells: Schneider's/ 10% FBS/ PS450 ml Schneider's medium (Gibco #11720-034)(pour off 50 ml into Falcon to save as serum-free~undefined*50_ml_Fetal_Bovine__...

This is our modification of a recipe used by John Loera at the University of Texas at Austin. The Texas recipe is apparently based on the malt medium described by Lakovaara (1969) with the addition ...

Cuticle preparationsThis procedure was adapted from one described over the phone (thanks P.G.). We're not sure if it appears elsewhere in published form.-Collect embryos on apple juice-agar plates for ...

GENERAL MAINTENANCE INSTRUCTIONS FOR IMAGINAL DISC CELL LINES This is our lab protocol for growing imaginal disc cells (clone-8) The Milner Lab website has additional protocols for clone-8 cells. Cl ...

This is a firm medium that keeps well and makes a good base for egg collecting. Wrap filled petri dishes and store at 4-- 14oC (they should keep for at least 6 weeks). Substitute molasses for the c ...

Sporulation in Liquid Media1) Grow cells in 25mls of PSP2 (in 250ml flask) supplemented with 25% of the recommended concentration of amino acids until they are about 1x107 cells/ml (~1 day).2) Wash wi ...

YPD FILLED MICROTITER PLATES Uses: 1. Grow individual strains to use for PCR2. Shipping strains 1. Make YPD regular agar (1.5-2%) for sending strains.2. Make 200ml batches = enough for 6 microtiter pl ...

HC Dropout Plate Supplementation TableAmino AcidStockAmount To Spread(per 25 ml plate)URA1 mg/ml875 µlADE1 mg/ml500 µlLEU20 mg/ml100 µlLYS10 mg/ml300 µlMET2 mg/ml250 µlHIS10 mg/ml50 µlTRP10 mg/ml200 µ ...

Fly extract 2 for routine culture (FE2) 1. First collect your flies; tip them from culture bottles into an empty glass bottle and plug it. Put it in the freezer for 45 mins or so until the flies are ...

Production of Conditioned Medium containing Wingless protein from S2 cells- Grow heatshock wingless S2 cells at 25 degrees centigrade in a tissue culture flask to approximately 2.5 x 106 cells/ml.- He ...

Medium Shields and Sang Medium 3 (SS3) modified for low serum. Supplemented before use with 2% heat inactivated FCS 2.5% FE2 12.5IU/100ml insulin and filter-sterilised through 0.22µm filter. Supplemen ...