Friday, December 19, 2003 Description This assay uses the zigmond chamber to study chemotaxis of spermatozoa or other migratory cells. The chamber consists of two wells separated by a bridge. The culture medium is placed on the bridge, cov...

Thursday, December 04, 2003 Description After extraction of soluble protein from leaves, samples are run on a SDS-PAGE gel. The method according to Sambrook et al. (1989) is used for SDS-PAGE and the method as described in the technical bo...

Description The specific cysteine proteinase substrate Z-Phe-Arg-MCA is used as described by Barret and Kirschke (1981). The substrate, Z-Phe-Arg-NMec, which is barely fluorescent, is hydrolyzed to liberate 7-amino-4-methylcoumarin, and th...

Thursday, December 04, 2003 Description The histochemical assay to screen for expression of b-glucurodinase (gus) activity in genetically modified plants is carried out according to the method of Jefferson et al. (1987) Procedure For analy...

Thursday, December 04, 2003 Description Transient Expression Assay Using Maize Mesophyll Protoplasts Procedure Plant Growth Soak seeds in water for overnight. Use half vermiculite and half pelite moss and germinate seeds under light for 3...

Thursday, December 04, 2003 Description Transient Expression Assay Using Arabidopsis Mesophyll Protoplasts : PEG Transfection Procedure All steps are carried out at 23oC Add 20 ml DNA (20-40 mg of plasmid DNA of 5 kb in size) Add 200 ml pr...

Thursday, December 04, 2003 Description Transient Expression Assay Using Arabidopsis Mesophyll Protoplasts : Protoplast Isolation Procedure Plant Materials: BE plants grown on the B5 medium Greenhouse-grown BE, Col, Ler and C24 plants are...

Thursday, December 04, 2003 Description PP2C assay Procedure 1. Phosphorylation of casein Casein kinaseI (NEB), 1000U/ul Sigma casein C-4765, 5% solution (56mg/ml), dilute to 10mg/ml 500ul reaction, 30oC, 4h 50 mM Tris-HCl, pH 7.5 10 mM Mg...

Description Rice Bombardment Procedure DAY 1 1. Filtering Rice cells with 640 m mesh to get small size cells. 2. Discarding the sup 3. Mountaining #4 whatman paper on the N6D 0.5 media 4. Spread 250mg cell on the paper circulary, 1-layerd....

Description Agrobacterium-mediated Tobacco Transformation Procedure 1. Agrobacterium culture and Harvest: Agrobacterium overnight culture in YEP cfg 5000pm, washing in MS, dilution (1/10 of original volum) with MS 2. Tobacco leaf steriliza...