其他模型

Antibody Staining of Dissected Gonads - From R. Francis - -Adapted by Min-Ho Lee-Antibody staining (in Tubes) 1. Using a pasteur pipette (drawn in flame and cut open) transfer worm carcasses with a ...

RUVKUN Antibody Staining Protocol revised 10/29/90Fixation: 1. C. elegans N2 grown in liquid or plates. Harvest worms.Wash for 2 hours at room temp. in 1X M9 or PBS. 2. Fix in 5 ml of FRESH THAT DAY 1 ...

C. elegans whole mount immunohistochemistry with Bouin's FixativeI have developed a fixation protocol which provides an alternative set of conditions to test antisera that work poorly on paraformaldeh ...

DAPI Staining To visualize DNA incubate fixed samples with 100 ng/ml 4'6'-diamidino-2-phenylindole hydrochloride (DAPI) in PBS for 30 min. Rinse 3x with PBTw. MaterialsPBS: Sambrook et al. (Molecular ...

Double dye filling (using DiO and di-4 ANEPPS)(Krista Williams) Dye Filling Label 15 ml centrifuge (c/f) tubes with strain name. Squirt ~1-2 ml M9 onto plate with a Pasteur pipet. Rinse plate with M9 ...

Dye Filling to Stain Amphid and Phasmid Neurons by Michael Koelle from Beth Sawin 4/6/94 1. Buy DiO from Molecular Probes catalog # D-275. DiO fluorescesces green (use FITC filters). If you want red f ...

Fixation and permeabilization protocol 5/30/91 Mike FinneyThis protocol works for all stages except dauers (which won't open) andhypoclorite-treated eggs (which disintegrate). Luckily hypochloritetrea ...

Gonad Dissections- From R. Francis -1) Pick adults and/or L4s to an unseeded NGM plate. Alternatively wash worms off a seeded plate with 1.5 ml of PBS and allow to gravity settle for a few minutes in ...

Gonad Fixation - From R. Francis -A. Methanol Methanol fixation is quick easy and gives satisfactory nuclear morphology after DAPI staining. However methanol-fixed gonads break easily and are more dif ...

Gentle hypochlorite treatment1. Wash off plates of gravid hermaphrodites with 10-15 mls of M9 and put in 15 ml conical tube. Collect minimum of 4 or 5 plates in a tube.2. Centrifuge @500rpm for about ...

In situ hybridizationIn situ hybridizations were based on a modified method of Seydoux and Fire (1994 Development 120 2823-2834; 1995 Methods in Cell Biology) (H. Tabara and Y. Kohara personal commun ...

MMS integration (both methods modified by Herman Lab) NOTES: Remember that MMS is a potent mutagen -- wear gloves and be very careful. Integration is best done using an array that is transmitted at ve ...

Nematode dye filling(Ed Hedgecock ) Thumb plate Transfer worms into a known amount of M9 in a labeled depression. Add DiO(2 mg/ml in DMF) at 10 l/ml of M9. Let sit for 2 hours. Transfer worms with a P ...

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 ...