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Preparation and Fractionation of Drosophila Embryo Extracts for the In Vitro Characterization of RNA - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4324

Category: RNAi Technology Protocols: RNAi Drosophila Protocols

Protocol describes a method to prepare and fractionate Drosophila embryo extracts. These extracts can be used for the in vitro characterization of RNAi. - [Read Preparation and Fractionation of Drosophila Embryo Extracts for the In Vitro Characterization of RNA]

Preparation and High-efficiency Transformation of Competent E. coli - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3942

Category: Bacteria Transformation Protocols

Protocol generates competent cultures of E. coli that can be transformed at high frequencies (5 x 108 transformed colonies/µg of superhelical plasmid DNA). - [Read Preparation and High-efficiency Transformation of Competent E. coli]

Preparation and LCM of Paraffin Embedded Tissue Sections Protocol - http://dir.nichd.nih.gov/lcm/LCMTAP.htm#II.%20Preparation%20and%20LCM%20of%20Paraffin%20Embedded%20Tissue%20Sections

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

Protocol for the preparation of LCM of Paraffin embedded tissue sections. Includes: Fixation; Processing, Embedding and Tissue Sectioning; Staining; Laser Transfer; DNA Extraction Protocol of LCM tissue and PCR for DNA analysis; RNA Extraction Protocol of LCM tissue and RT-PCR for mRNA analysis. - [Read Preparation and LCM of Paraffin Embedded Tissue Sections Protocol]

Preparation and Loading of Protein Samples for Microinjection Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4656

Category: Microinjection Protocols: Microinjection of Proteins Protocols

his protocol provides methods for the preparation of protein samples and for loading them into pulled microinjection pipettes. Stock solutions of proteins are thawed, diluted (if desired), centrifuged at high speed to remove aggregates, and kept on ice until loading. Loading into micropipettes can be done using either a "front-loading" or a "backfilling" procedure. - [Read Preparation and Loading of Protein Samples for Microinjection Protocol]

Preparation and Microinjection of x-Rhodamine-Labeled Vimentin Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4676

Category: Cytoskeleton Protocols: Intermediate Filaments Protocols

Intermediate filaments (IF) are major cytoskeletal systems of vertebrate and many nonvertebrate cells whose expression is cell-type specific and developmentally regulated. This protocol describes the x-rhodamine labeling of one type of IF, vimentin, and a method for microinjection of the labeled vimentin into cultured cells. IF dynamics can then be examined with fluorescence microscopy. - [Read Preparation and Microinjection of x-Rhodamine-Labeled Vimentin Protocol]

Preparation and Transformation of Competent E. coli Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3942

Category: E Coli Protocols: E coli Transformation Protocols

Procedure generates competent cultures of E. coli that can be transformed at high frequencies (5 x 108 transformed colonies/µg of superhelical plasmid DNA). IMPORTANT All steps in this protocol should be carried out aseptically. - [Read Preparation and Transformation of Competent E. coli Protocol]

Preparation and Transformation of Competent E. coli Using Calcium Chloride Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3932

Category: Bacteria Transformation Protocols

Protocol used to prepare batches of competent bacteria that yield 5 x 106 to 2 x 107 transformed colonies/µg of supercoiled plasmid DNA. - [Read Preparation and Transformation of Competent E. coli Using Calcium Chloride Protocol]

Preparation and Transformation of Competent E. coli Using Calcium Chloride Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3932

Category: E Coli Protocols: E coli Transformation Protocols

Protocol is is used to prepare batches of competent bacteria that yield 5 x 106 to 2 x 107 transformed colonies/µg of supercoiled plasmid DNA. - [Read Preparation and Transformation of Competent E. coli Using Calcium Chloride Protocol]

Preparation and Transformation of Competent E. Coli: "Ultra-Competent" Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3944

Category: Bacteria Transformation Protocols

Protocol reproducibly generates competent cultures of E. coli that yield 1 x 108 to 3 x 108 transformed colonies/µg of plasmid DNA. The protocol works optimally when the bacterial culture is grown at 18°C. If a suitable incubator is not available, a standard bacterial shaker can be set up in a 4°C cold room and regulated to 18°C. - [Read Preparation and Transformation of Competent E. Coli: "Ultra-Competent" Cells Protocol]

Preparation and Transformation of Competent E. Coli: "Ultra-Competent" Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3944

Category: E Coli Protocols: E coli Transformation Protocols

Preparation of a Denaturing Agarose Gel for the Analysis of RNA Samples Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/14/pdb.prot4127

Category: Nucleic Acid Electrophoresis Protocols: RNA Electrophoresis Protocols

Protocol for preparation of a denaturing agarose gel for the analysis of RNA samples. The integrity of RNA samples can be analyzed by electrophoresis on a denaturing agarose gel containing formaldehyde. - [Read Preparation of a Denaturing Agarose Gel for the Analysis of RNA Samples Protocol]

Preparation of a Highly Efficient Transcription Extract from Drosophila Embryos Protocol - http://www.bio.com/protocolstools/protocol.jhtml%3Bjsessionid%3D3APITJJIWFIM1R3FQLMCFEWHUWBNSIV0?id=p9052

Category: Drosophila Protocols: Drosophila Genetics Protocols

Protocol describes how to produce a soluble nuclear extract rich in basal pol II transcription factors from Drosophila embryos. This is a cell-free extract that contains all the necessary transcription factors and is capable of accurate initiation of transcription by RNA polymerase II but is deficient in core histones and histone H1. - [Read Preparation of a Highly Efficient Transcription Extract from Drosophila Embryos Protocol]

Preparation of a Prepacked IMAC Column Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4214

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Protocol for the preparation of a prepacked IMAC column. Purification of histidine-tagged proteins on a milligram scale can be performed conveniently on a small (1to 5-ml) immobilized metal-ion chromatography (IMAC) column using a syringe to load the sample, wash the column, and elute the protein. - [Read Preparation of a Prepacked IMAC Column Protocol]

Preparation of Affinity-Ligand Resins by Immobilization of Dyes on Polyhydroxyl Matrices - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4208

Category: Chromatography Protocols: Dye Ligand Affinity Chromatography Protocols

Triazine dyes, such as Cibacron Blue 3GA, can be linked to a hexyl spacer arm and then immobilized on a polyhydroxyl support matrix that has been activated with either 1,1-carbonyldiimidazole or epichlorohydrin. An alternative procedure for immobilizing dyes using the direct coupling method is provided in Immobilization of Dyes on Polyhydroxyl Matrices Using the Direct Coupling Method. - [Read Preparation of Affinity-Ligand Resins by Immobilization of Dyes on Polyhydroxyl Matrices]

Preparation of Affinity-Ligand Resins by Immobilization of Dyes on Polyhydroxyl Matrices Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4207

Category: Chromatography Protocols: Dye Ligand Affinity Chromatography Protocols

Protocol describes a general approach for the direct attachment of a triazine dye onto a carbohydrate support. The direct coupling method is achieved under alkaline conditions (pH 10-11) with nucleophilic displacement of the dye’s chlorine atom by the polymer hydroxyls. - [Read Preparation of Affinity-Ligand Resins by Immobilization of Dyes on Polyhydroxyl Matrices Protocol]

Preparation of an Ion-Exchange Column Protocol - http://cshprotocols.cshlp.org/cgi/content/extract/2006/1/pdb.prot4195

Category: Chromatography Protocols: Ion Exchange Chromatography Protocols

Protocol for the preparation of ion-exchange chromatography column. Ion-exchange chromatography (IEC) can be used as a crude step in a protein purification scheme, or, with proper preparation, as a high-resolution step. If high resolution is desired, considerable care should be taken during column preparation, choice of IEC media, and column packing. - [Read Preparation of an Ion-Exchange Column Protocol]

Preparation of Arabidopsis Chloroplasts Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4681

Category: Plant Biology Protocols: Arabidopsis Tissue Culture

It is desirable to prepare subcellular fractions, either to localize proteins or to improve the sensitivity of protein detection. This procedure describes the enrichment of chloroplasts from Arabidopsis. - [Read Preparation of Arabidopsis Chloroplasts Protocol]

Preparation of Arabidopsis Mitochondria Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4682

Category: Plant Biology Protocols: Arabidopsis Tissue Culture

It is often desirable to prepare subcellular fractions, either to localize proteins or to improve the sensitivity of protein detection. This procedure describes the enrichment of mitochondria from Arabidopsis. - [Read Preparation of Arabidopsis Mitochondria Protocol]

Preparation of B-Lymphocyte Lysates for Cyclic AMP Determination - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000012.pdf?pid=PP00000012

Category: Signalling Signal Transduction Protocols

This protocol provides a sufficient sample for several determinations of cAMP using the acetylation protocol. The method chosen for measuring the content of cyclic adenosine 3',5'-monophosphate (cyclic AMP or cAMP) in splenic B lymphocytes (B cells) is an enzyme linked immunoassay system. Protocol includes information on: Treatment of Cells and Preparation of Extracts; Reagents and Materials. - [Read Preparation of B-Lymphocyte Lysates for Cyclic AMP Determination]

Preparation of B-Lymphocyte Lysates for Cyclic AMP Determination for Dual Ligand Screen - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000139.pdf?pid=PP00000139

Category: Signalling Signal Transduction Protocols

Protocol provides a method of achieving a sufficient sample for several determinations of cAMP using Amershamâ€™s acetylation protocol. Protocol includes information on: Treatment of Cells and Preparation of Extracts; Reagents and Materials. - [Read Preparation of B-Lymphocyte Lysates for Cyclic AMP Determination for Dual Ligand Screen]

Preparation of B-Lymphocyte Lysates for Cyclic AMP Determination for Dual Ligand Screen Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000139.pdf?pid=PP00000139

Category: Immunology: B Cell Protocols

The method chosen for measuring the content of cyclic adenosine 3',5'- monophosphate (cyclic AMP or cAMP) in splenic B lymphocytes (B cells) is an enzymelinked immunoassay system. Includes: Treatment of Cells and Preparation of Extracts. - [Read Preparation of B-Lymphocyte Lysates for Cyclic AMP Determination for Dual Ligand Screen Protocol]

Preparation of B-Lymphocyte RNA for Microarray Analysis Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000009.pdf?pid=PP00000009

Category: Immunology: B Cell Protocols

Procedure permits the isolation of at least 5 µg of total RNA from a sample of purified mouse splenic B lymphocytes. The quality of the RNA is assessed by separation of an aliquot through 1% agarose and staining with ethidium bromide as described in AfCS protocol Visualization of RNA Preparations on 1% Agarose Gels. The isolated RNA is used for analysis of gene expression by microarray technology. analysis of gene expression by microarray technology. - [Read Preparation of B-Lymphocyte RNA for Microarray Analysis Protocol]

Preparation of B-Lymphocyte RNA for Microarray Analysis Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000009.pdf?pid=PP00000009

Category: Microarray Protocols: Microarray Analysis Protocols

Protocol permits the isolation of at least 5 µg of total RNA from a sample of purified mouse splenic B lymphocytes Isolation of Resting B Lymphocytes from Sixteen Mouse Spleens. - [Read Preparation of B-Lymphocyte RNA for Microarray Analysis Protocol]

Preparation of BAC DNA with CONCERT High Purity Plasmid Purification System Protocol - http://intramural.nimh.nih.gov/lcmr/snge/Protocols/BACProtocols/BACColumn.html

Category: BACs Bacterial Artificial Chromosome Protocols

Protocol for the preparation of BAC (Bacterial Artificial Chromosome) DNA with CONCERT high purity plasmid purification system. Includes: - [Read Preparation of BAC DNA with CONCERT High Purity Plasmid Purification System Protocol]

Preparation of Bacterial Glycerol Stocks, Thawing of Stocks - http://sbio.uct.ac.za/Sbio/documentation/Glycerol%20stocks.html

Category: Buffers Media, and Solutions: Bacterial Solutions

Preparation of Bacterial Glycerol Stocks, Thawing of Stocks. South African Structural Biology Initiative. - [Read Preparation of Bacterial Glycerol Stocks, Thawing of Stocks]

Articles (1-10 of 13)

Preparing Stock Plating of Bacteria Protocol

A protocol for the preparation of stock plating of bacteria.

Lambda Phage Protocol for Large Preparation DNA Recovery

Lambda Phage Protocol for Large Preparations DNA Recovery.

Lambda Phage Protocol Large DNA Preparation

Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA

This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.

Probe Preparation for 22 x 40 mm DNA Microarrays

Probe Preparation for 22 x 40 mm DNA Microarrays Protocol.

Microinjection Pipettes Protocol

Microinjection Pipettes Preparation Protocol.

In Vitro Translated Xenopus Mos Kinase Assay Protocol

In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.

Immobilized Antigen Phage Antibody Selection Protocol

A protocol for the selection of Phage Antibodies using Immobilized Antigen. This method describes the selection of antibodies from bacteriophage antibody libraries that recognize a specific antigen. The phage display library of antibody-displaying phage particles is exposed to antigen attached to a solid substrate (Nunc Immuno™ tubes). The phage particles with affinity for antigen bind to the immobilized antigen and are selected from the library of phage expressing antibodies.