Description This protocol produces RNA transcripts from a cloned gene that can then be used in an in vitro translation assay. Procedure 1. Digest the gene-containing plasmid DNA to completion with a restriction endonuclease to linearize th...

Description This protocol describes an RNA Polymerase III (Pol III) transcription assay using an extract or proteins of choice. Pol III is the polymerase responsible for transcribing 5S RNA, tRNAs, and other small RNAs. á-Amanitin inhibit...

Description This protocol describes how to perform a DEAE-dextran transfection of COS cells and provides information on labeling the expressed protein with [35S-Cysteine] for SDS-PAGE analysis. Procedure A. Transfection of COS-7 Cells with...

Sunday, October 19, 2003 Description This protocol can be used to measure the amount of incorporation of 35S-Cysteine into cellular protein following a metabolic labeling procedure. This protocol reduces the amount of non-specific counts,...

Saturday, October 18, 2003 Description This procedure allows detection of specific cell adhesion to glycolipids resolved on TLC plates. The steps include: (1) HPTLC of glycoconjugates, (2) coating the chromatogram with polymer, (3) prebloc...

Saturday, October 18, 2003 Description Matirgel is considered as basement membrane and generated from EHS sarcoma. Matrigel contains not only basement membrane components (collagens, laminin, and proteoglycans)but also matrix degrading enz...

Saturday, October 18, 2003 Description A modification of the basic immunofluorescence staining and flow cytometric analysis protocol can be used for the simultaneous analysis of surface molecules and intracellular cytokines at single-cell...

Friday, October 17, 2003 Description H-Thymidine Uptake by Cultured Cells Procedure Grow either L cells (Mouse fibroblasts) or chick embryo fibroblasts on coverglasses and then give them H-thymidine for a short period of time. At the end o...

Thursday, October 16, 2003 Description A 2 days program to obtain tranformant Listeria Procedure 1 - Preparation of competent Listeria monocytogenes Day 1: 1.Prepare reagents needed: - 2X BHI (37g BHI for a final volume of 500ml, autoclave...

Thursday, October 16, 2003 Description Chromosome preparation and Fluorescence in-situ hybridization (FISH) Procedure Y To be read as micro (mu) 1.Thin slides were kept in chromic acid overnight. 2.The slides were rinsed in running tap wat...