embl Protocols

Category: siRNA and RNAi Protocols

DSRNAI EXPERIMENTS IN DROSOPHILA S2 CELL CULTURE. using T7 to create RNA. Carolyn Worby. Dixon Lab RNAi experiments. - [Read DSRNAI EXPERIMENTS IN DROSOPHILA S2 CELL CULTURE]

Category: DNA Protocols: DNA Cloning

Preparation of oligo solutions PCR experiments, Digestion of insert DNA, Digestion and dephosphorylation of vector DNA, Ligation of DNA fragments with sticky ends, Ligation of DNA fragments with blunt ends, Preparation of chemically competent E. coli - [Read EMBL Cloning protocols]

Category: Proteomic Protocols

In Gel Digestion Protocols for Silver Stained and Coomassie Stained Gels. (EMBL protocol). - [Read In Gel Digestion Protocols for Silver Stained and Coomassie Stained Gels]

Category: Proteomic Protocols: 2-D Gel Electrophoresis

In gel trypsin digestion for Mass spectrometry. (According to EMBL Protein and Peptide Group, 1997) - [Read In-Gel Digestion of Proteins Separated by Polyacrylamide Gel Electrophoresis]

Category: Proteomic Protocols

In-Gel Digestion Protocol. Excision of protein bands (spots) from polyacrylamide gels Reduction and alkylation . Reduction and alkylation . MALDI analysis of the supernatant after in-gel digestion. Extraction of Peptides.Matthias Wilm EMBL Bioanalytical R - [Read In-gel digestion of proteins to be analyzed by mass spectrometry]

Category: Mass Spectrometry Protocols

MALDI Matrices. Commonly used MALDI matrices for analysis of peptides, proteins, carbohydrates, and nucleic acids using 337 nm or 355 nm UV lasers. All matrices can be used for sample preparation using the Dried Droplet Method whereas only matrices solubl - [Read MALDI Matrices]

Category: Mass Spectrometry Protocols

Method for sample preparation for MALDI: dried droplet protocol. EMBL Bioanalytical Research Group. Matthias Wilm . - [Read MALDI Sample Preparation - Dried Droplet Method]

Category: Mass Spectrometry Protocols

Fast Evaporation Method for MALDI sample preparation protocol. Inserting into MALDI mass spectrometer and acquiring a spectrum. EMBL Bioanalytical Research Group. Wilm. - [Read MALDI Sample Preparation - Fast Evaporation Method]

Category: Mass Spectrometry Protocols

Mass spectrometry compatible silver staining of gels protocol. EMBL Bioanalytical Research Group. Wilm. - [Read Mass spectrometry compatible silver staining of gels protocol]

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Bacteriophage {lambda} vectors that allow genetic selection of recombinant bacteriophages (e.g., the EMBL series, {lambda}2001, {lambda}DASH, {lambda}ZAP, and {lambda}gt10) can be prepared for cloning by simple digestion by one or more restriction enzymes. - [Read Preparation of Bacteriophage lambda DNA Cleaved with a Single Restriction Enzyme Protocol]

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Many replacement vectors (e.g., the EMBL series, {lambda}2001, and {lambda}DASH) contain a series of restriction sites, arranged in opposite orientations, at each end of the central stuffer fragment. Digestion of these vectors with two different restriction enzymes yields left and right arms, a stuffer fragment, and short segments of the polycloning sites. These can easily be removed from the arms by differential precipitation with isopropanol or spun-column chromatography. - [Read Preparation of Bacteriophage lambda DNA Cleaved with Two Restriction Enzymes Protocol]

Category: Protein Protocols: Protein Extraction From Cells

Preparation of cell lysates from yeast by enzymatic digestion of the cell wall. - [Read Preparation of cell lysates from yeast by enzymatic digestion of the cell wall]

Category: Protein Protocols: Protein Extraction From Cells

Preparation of cell lysates from yeast using a French Press. he Protein Expression and Purification Facility, EMBL - [Read Preparation of cell lysates from yeast using a French Press]

Category: Protein Protocols: Protein Extraction From Cells

Preparation of cell lysates from yeast using glass beads vortexing. Includes lysis buffer recipes. The Protein Expression and Purification Facility, EMBL - [Read Preparation of cell lysates from yeast using glass beads vortexing]

Category: Protein Protocols: Protein Expression Protocols

Protein Expression and Purification Flowchart. EMBL - [Read Protein Expression and Purification Flowchart]

Category: siRNA and RNAi Protocols

RNA interference protocol. Preparation of Template DNA, Preparation of the dsRNA, RNAi in Drosophila Schneider cells. Matthias Wilm. - [Read RNA interference protocol]

Category: Genetics and Genomics: Cancer Genetics Protocols

SAGE is a new method that has been invented at Johns Hopkins University in USA to give scientists an overview of a cell’s complete gene activity. It works by capturing RNAs, identifying them and counting them. By comparing different types of cells, the researchers hope to generate profiles that will help them understand healthy cells and what goes wrong during diseases. Includes: How SAGE works and Steps of SAGE. - [Read Serial Analysis Of Gene Expression (SAGE)]