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Preparation of Segmented and Polarity Marked Microtubules Protocol - http://mitchison.med.harvard.edu/protocols/polarity.html

Category: Cytoskeleton Protocols: Microtubules Protocols

Segmented and polarity-marked microtubules are very useful for many different types of in vitro assays. Segmented microtubules are microtubules with a bright seed and dim elongated segments on both ends. Polarity marked microtubules are microtubules with a bright seed and a dim elongated segment only on one end -- the plus end. - [Read Preparation of Segmented and Polarity Marked Microtubules Protocol]

Preparation of Single-stranded Bacteriophage M13 DNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3686

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

Bacteriophage M13 single-stranded DNA is prepared from virus particles secreted by infected cells into the surrounding medium. The filamentous particles are concentrated by precipitation from a high-ionic-strength buffer with polyethylene glycol. Subsequent extraction with phenol releases the single-stranded DNA, which is then collected by precipitation with ethanol. This protocol is generally used to prepare single-stranded DNA from a small number of M13 isolates. - [Read Preparation of Single-stranded Bacteriophage M13 DNA Protocol]

Preparation of siRNAs from Drosophila Embryo Extracts Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4325

Category: RNAi Technology Protocols: RNAi Drosophila Protocols

siRNAs produced upon the addition of dsRNA to Drosophila embryo extract are enriched in a micrococcus-nuclease-resistant fraction. After proteinase K treatment and dephosphorylation with calf intestinal phosphatase, these siRNAs mediate efficient RNAi in vitro. - [Read Preparation of siRNAs from Drosophila Embryo Extracts Protocol]

Preparation of Solid Tissue for Aspergillus Galactomannan Antigen Detection by Platelia Protocol - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./galactoman.html~main

Category: Fungi Protocols

Protocol for the preparation of solid tissue for Aspergillus galactomannan antigen detection by Platelia (Biorad). Technique was designed for use on human serum. However, it may also be possible to perform this method on solid tissues and organic solutions. Viscous solution and tissue specimens need to be pre-treated to achieve the extraction of the Aspergillus antigen and to get a homogeneous sample in solution. - [Read Preparation of Solid Tissue for Aspergillus Galactomannan Antigen Detection by Platelia Protocol]

Preparation of Stock Solutions for FISH Protocol - http://users.ugent.be/~fspelema/neubla/protocol/stock.htm

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Protocol for the preparation of stock solutions for FISH. - [Read Preparation of Stock Solutions for FISH Protocol]

Preparation of the 32P-Labeled Oligonucleotide Probe Protocol - http://www.biochem.northwestern.edu/ibis/morimoto/research/Protocols/III.%20Proteins/C.%20DNA-Protein%20Interactions/4.%2032P-label.pdf

Category: DNA Protein Interactions Protocols

Protocol for preparation of the 32P-labeled oligonucleotide probe. - [Read Preparation of the 32P-Labeled Oligonucleotide Probe Protocol]

Preparation of ultra-competent E. coli cells for transformation - http://www.hybtech.org/Liu/uccell.html

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Preparation of ultra-competent E. coli cells for transformation. Dr. Chun-Ming Liu. Based on Inoue et al (1990). - [Read Preparation of ultra-competent E. coli cells for transformation]

Preparation of Uracil-containing Single-stranded Bacteriophage M13 DNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3811

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

Single-stranded templates of bacteriophage M13 DNA containing 20-30 residues of uracil in place of thymine are generated during growth of the bacteriophage in an F' strain of E. coli carrying mutations in the ung and dut genes. This DNA is used as a template in the Kunkel method of oligonucleotide-directed mutagenesis (Oligonucleotide-directed Mutagenesis of Single-stranded DNA). - [Read Preparation of Uracil-containing Single-stranded Bacteriophage M13 DNA Protocol]

Preparation of whole cell homogenates of cultured mammalian cells for 2-D Electrophoresis - http://www.healthsystem.virginia.edu/internet/biomolec/2dgelsampleprep.cfm

Category: Proteomic Protocols

Preparation of whole cell homogenates of cultured mammalian cells for 2-D Electrophoresis. Univ. Virginia. - [Read Preparation of whole cell homogenates of cultured mammalian cells for 2-D Electrophoresis]

Preparation of Worm Extracts by Sonication Protocol - http://www.genetics.wustl.edu/tslab/Protocols/Cyto_extract_sonicat_v2.htm

Category: C. elegans Protocols

Protocol was designed to rapidly generate small scale cytosolic extracts of C. elegans for Western or IP (has not been tested for RNA work). The protocol works well for between 50 to 5000 worms and has not been extensively tested on larger a scale, though it should work. Includes: Collection; Sonication; Clearing lysate; Immunoprecipitation. - [Read Preparation of Worm Extracts by Sonication Protocol]

Preparation of Xenopus Sperm Nuclei Protocol - http://www.bio.com/protocolstools/protocol.jhtml?id=p877

Category: Xenopus Protocols: Xenopus Egg Extracts Protocols

Protocol describes the preparation of nuclei from Xenopus sperm. These nuclei can be used for spindle assembly assays, microscopy, or DNA synthesis. - [Read Preparation of Xenopus Sperm Nuclei Protocol]

Preparation of Yeast Cells for Flow Cytometry Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4178

Category: Yeast Protocols: Yeast Nucleic Acid Protocols

Flow cytometry is used to analyze the quantity of DNA in cells. Since the DNA content of cells varies through the cell cycle, this information can provide an indication of cell cycle progression. This protocol uses SYTOX Green staining. - [Read Preparation of Yeast Cells for Flow Cytometry Protocol]

Preparation of Yeast DNA Embedded in Agarose Plugs Protocol - http://fangman-brewer.genetics.washington.edu/plug.html

Category: Yeast Protocols: Yeast Nucleic Acid Protocols: Yeast DNA Isolation Protocols

Protocol for preparation of Yeast DNA embedded in agarose plugs. - [Read Preparation of Yeast DNA Embedded in Agarose Plugs Protocol]

Preparation, LCM and RNA/DNA extraction of Frozen Tissue Sections Protocol - http://dir.nichd.nih.gov/lcm/LCMTAP.htm#I.%20Preparation,%20LCM%20and%20RNA/DNA%20extraction%20of%20Frozen%20Tissue%20Sections

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

Protocol for the preparation, LCM and RNA/DNA extraction of frozen tissue sections. Includes: Embedding, Cutting and Staining. - [Read Preparation, LCM and RNA/DNA extraction of Frozen Tissue Sections Protocol]

Preparing a Cell Block from Cell Lines Cell Suspensions Protocol - http://icg.cpmc.columbia.edu/cattoretti/Protocol/immunohistochemistry/Cell_block.html

Category: Immunohistochemistry Protocols: Blocking Protocols

Protocol for preparing a cell block from cell lines / cell suspensions. Includes: Live cell preparation; Fixed cell preparation. - [Read Preparing a Cell Block from Cell Lines Cell Suspensions Protocol]

Preparing Chemically Competent Cells - http://openwetware.org/wiki/Preparing_chemically_competent_cells

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Detailed protocol Preparation and Steps for E.Coli Competent Cells procedure. Open Net Ware. - [Read Preparing Chemically Competent Cells]

Preparing Denatured Templates for Sequencing by Dideoxy-mediated Chain Termination Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3787

Category: DNA Protocols: DNA Sequencing Protocols: Dideoxy Mediated Sequencing Protocols

Protocol describes how double-stranded plasmid DNA is denatured byalkali and annealed to an appropriate oligonucleotide primer,in preparation for dideoxy-sequencing reactions catalyzed bySequenase. - [Read Preparing Denatured Templates for Sequencing by Dideoxy-mediated Chain Termination Protocol]

Preparing Embryonic Stem (ES) Cells for Aggregation Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4419

Category: Stem Cell Protocols

Ths protocol describes the preparation of ES cells, which can then be aggregated with diploid or tetraploid embryos to produce aggregation chimeras. - [Read Preparing Embryonic Stem (ES) Cells for Aggregation Protocol]

Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4400

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the preparation of feeder cells from MEF cells or from the STO mouse fibroblast cell line. The cells are rendered mitotically inactive by treatment with {gamma}-irradiation. The feeder layers can then be used to maintain embryonic stem (ES) cells in the undifferentiated state. - [Read Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells Protocol]

Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells: Treatment with Mitomyc - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4399

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the preparation of feeder cell layers from MEF cells or from the STO mouse fibroblast cell line. The cells are rendered mitotically inactive by treatment with mitomycin C. The feeder layers can then be used to maintain embryonic stem (ES) cells in the undifferentiated state. - [Read Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells: Treatment with Mitomyc]

Preparing Mouse Embryo Fibroblasts Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4398

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the preparation of mouse embryo fibroblasts (MEFs), which can then be used as feeder cells to maintain embryonic stem (ES) cells in the undifferentiated state. - [Read Preparing Mouse Embryo Fibroblasts Protocol]

Preparing the Aggregation Plate Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4420

Category: Mouse Protocols: Assembling Chimeras Mouse Protocols

Protocol describes the preparation of aggregation plates, which will then be used for the production of aggregation chimeras. - [Read Preparing the Aggregation Plate Protocol]

Principles and Applications of Liquid Scintillation Counting PDF - http://www.ehs.psu.edu/radprot/LSC_Theory2.pdf

Category: Radioactivity: Liquid Scintillation Counting

Excellent guide for Liquid Scintillation Counting. Includes protocols and methods for counting gel slices, SPECIAL SAMPLE PREPARATION PROTOCOLS: TLC Plates, protocol for Counting Samples on Cellulose-ester, Filters (MilliporeTM filters), Counting Tissue radioacitivity, Counting 14CO2, Samples in Polyacrylamide Gels. National Diagnostics. National Diagnostics Laboratory Staff. - [Read Principles and Applications of Liquid Scintillation Counting PDF]

Principles of Aseptic Technique: ESSENTIALS FOR ANIMAL RESEARCH - http://research.ucsb.edu/connect/acc/esasept.htm

Category: Animal Research Techniques

Facilities, equipment, sterilization, steam, gas, liquid, animal research, preparation of the animal, surgical pack, surgeon preparation. - [Read Principles of Aseptic Technique: ESSENTIALS FOR ANIMAL RESEARCH]

Probing Protein Interactions Using GFP and FRET Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3893

Category: Reporter Gene Assays: GFP Assay Protocols

In preparation for FLIM-FRET analysis, the appropriate donor and acceptor components must be introduced into live or fixed cells. The method of introduction depends on the nature of the components and the state of the cells. For example, plasmid DNAs encoding a protein of interest fused to a variant of GFP may be introduced into live cells by transfection or microinjection, whereas labeled antibodies are delivered by microinjection. - [Read Probing Protein Interactions Using GFP and FRET Protocol]

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.