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Amplification of RNA: High-Temperature Reverse Transcription and DNA Amplification with a Magnesium - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4115

Category: PCR Protocols: Reverse Transcriptase RT-PCR Protocols

Protocol uses a single thermostable RNA polymerase to perform high-specificity RT-PCR. A high-temperature RT reaction is followed by PCR amplification of the cDNA using a single thermostable poymerase, the GeneAmp AccRT RNA PCR enzyme from Applied Biosystems. The high temperature of the RT reaction enhances the specificity of primer binding and also reduces secondary structure in the template, thereby increasing the efficiency of polymerization. - [Read Amplification of RNA: High-Temperature Reverse Transcription and DNA Amplification with a Magnesium]

Biogenesis and Function of Peroxisomes and Glycosomes - http://students.washington.edu/mdminer/pabio_551/kessler1.pdf

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols: Peroxisome Isolation Protocols

Peroxisomes of higher eukaryotes, glycosomes of kinetoplastids, & glyoxysomes of plants are related microbody organelles that perform differing metabolic functions tailored to their cellular environments. The close evolutionary relationship of these organelles is most clearly evidenced by the conservation of proteins involved in matrix protein import and biogenesis. glycosome can be viewed as an offshoot of the peroxisomal lineage with additional metabolic functions, specifically glycolysi - [Read Biogenesis and Function of Peroxisomes and Glycosomes]

Cell Immortalization - http://www.atcc.org/common/products/CellImmortOverview.cfm

Category: Cell Immortalization Protocols

Information why cell immortalization is important. Also the available tools to be able to perform cell immortalization. - [Read Cell Immortalization]

Cotransfection of Luciferase Reporter Plasmids with siRNA Duplexes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4342

Category: Reporter Gene Assays: Luciferase Assay Protocols

It is possible that some cell lines have lost the ability to perform RNAi or that cells derived from certain tissues do not support RNAi. This reporter assay, for RNAi in mammalian cells, can be used to establish whether the cells under study are susceptible to RNAi. - [Read Cotransfection of Luciferase Reporter Plasmids with siRNA Duplexes Protocol]

Delivery of dsRNA into Drosophila Embryos by Gene Gun Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4517

Category: RNAi Technology Protocols: RNAi Drosophila Protocols

Protocol describes how subcellular-sized particles are accelerated to high velocity to carry double-stranded RNA (dsRNA) into Drosophila embryos. The major advantage of this procedure over microinjection (Microinjection of dsRNA into Drosophila Embryos) is that particle bombardment is easier and faster to perform. In addition, the mechanical trauma received is far less than by microinjection, allowing better survival of embryos and fewer phenotypic artifacts. - [Read Delivery of dsRNA into Drosophila Embryos by Gene Gun Protocol]

Immunostaining Tutorial - http://www.bcm.edu/rosenlab/protocols/IHC101.pdf

Category: Immunohistochemistry Protocols

Tutorial on immunostaining. Learn how to perform immunostaining for immunohistochemistry. - [Read Immunostaining Tutorial]

Manual Microdissection - http://cgap-mf.nih.gov/Protocols/Microdissection/ManualMicrodissection.html

Category: Primary Cell Isolation and Culture Protocols

There are several manual methods that can be used to perform tissue microdissection. Techniques using hand-held tools as well as mechanical micromanipulator-based approaches have been described. However, speed and precision are the most important parameters and any method that achieves these is adequate. Investigators should also expect to invest time initially by practicing on 10 to 20 cases to begin to feel comfortable with the technique. - [Read Manual Microdissection]

Molecular and Biochemical Analysis of Arabidopsis Protocol - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_197-207.pdf

Category: Plant Biology Protocols

The standard protocol for in situ hybridizations in plants still involves fixing fresh tissue, embedding the tissue in wax, sectioning with a microtome and detection of the transcripts of interest using labeled RNA-probes. This protocol concentrates only on nonradioactive methods, as they are easy to perform, very sensitive and even faster than techniques involving radioisotope labels. - [Read Molecular and Biochemical Analysis of Arabidopsis Protocol]

Monosaccharide Analysis by Methanolysis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4247

Category: Chromatography Protocols: Gas Chromatography Protocols

Protocol describes monosaccharide analysis by methanolysis. The method gives more complex chromatograms than the corresponding alditol acetate method, because up to four methyl glycosides can be produced from each monosaccharide. However, the method is easier to perform. - [Read Monosaccharide Analysis by Methanolysis Protocol]

Multiplexing Cell Viability Assays Protocols - http://www.promega.com/paguide/chap4.htm#title6

Category: Cell Biology and Cell Culture: Cell Viability Protocols

The latest generation of Promega cell-based assays includes luminescent and fluorescent chemistries to measure markers of cell viability, cytotoxicity and apoptosis, as well as to perform reporter analysis. Using these tools researchers can investigate how cells respond to growth factors, cytokines, hormones, mitogens, radiation, effectors, compound libraries and other signaling molecules. The protocols provided are guidelines for multiplexing cell-based assays & are intended as starting points. - [Read Multiplexing Cell Viability Assays Protocols]

PCR From TIssue Protocol - http://www.unizh.ch/botinst/Cyto_Website/schneitzLab/Methods/PCR/directTissuePCR.html/

Category: Plant Biology Protocols: Plant PCR

Protocol on how to perform a PCR from plant tissue (leaf or flower). - [Read PCR From TIssue Protocol]

Plant Crosses Protocol - http://www.unizh.ch/botinst/Cyto_Website/schneitzLab/Methods/PlantWork/crosses.html

Category: Plant Biology Protocols: Plant Genetics Protocols

Protocol describing on how to perform crosses in plants. - [Read Plant Crosses 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]

Protein Purification Using Immobilized Metal-Ion Affinity Chromatography under Optimized Imidazole - http://www.cshprotocols.org/cgi/content/extract/2006/9/pdb.prot4262

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Perform this protocol after determining the imidazole concentration that yields the best purification of the recombinant protein of interest. - [Read Protein Purification Using Immobilized Metal-Ion Affinity Chromatography under Optimized Imidazole]

Standard Chromatographic Conditions for RP-HPLC of Proteins Protocol - http://cshprotocols.cshlp.org/cgi/content/extract/2006/1/pdb.prot4200

Category: Chromatography Protocols: Liquid Chromatography Protocols

Protocol describes how to perform a test separation using a set of protein standards that have been designed to evaluate column performance. - [Read Standard Chromatographic Conditions for RP-HPLC of Proteins Protocol]

The Virtual Mouse Necropsy - http://www.geocities.com/virtualbiology/

Category: Animal Research Techniques

Perform a Mouse Necropsy on the Web. Dr. J.M.Ward - [Read The Virtual Mouse Necropsy]

Articles (1-3 of 3)

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.

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol. This protocol contains the steps for 3' end rapid amplification of mRNA by PCR. The first-strand cDNA is synthesized from total or poly(A+) RNA by priming from the poly-A tail of the mRNA using a oligo (dT) adaptor primer. The cDNA is then amplified via PCR using a gene-specific primer and an adaptor primer.

Vector Design Protocol for Transgenic Mice Generation

The protocol gives general considerations for the design of targeting vectors for transgenic mice. The protocol shares tips in the design of knock-out and knock-in vectors and some of their strategies for producing homologously recombined embryonic stem cells.

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