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Protocol: Purification of In Vitro Synthesized mRNA with Microcon or Centricon Centrifugal Filters - http://www.millipore.com/techpublications/tech1/6dlk4v

Category: Cell Biology and Cell Culture: In Vitro Reconstitution Protocols

In vitro transcription reactions employing T3, T7 or SP6 phage-encoded RNA polymerases are widely used to synthesize RNA from recombinant vectors containing appropriate promoters. Production of large amounts of specific RNA is valuable in the preparation of hybridization probes and in vitro translation studies; in the synthesis of ribozymes, rRNA, SRP, antisense RNA and substrates for RNA splicing; and in RNA-protein interaction studies. - [Read Protocol: Purification of In Vitro Synthesized mRNA with Microcon or Centricon Centrifugal Filters]

Protocols for LCM Preparation and Analysis - http://dir.nichd.nih.gov/lcm/LCMTAP.htm

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

Protocols for LCM preparation and analysis. Includes protocols: Preparation, LCM and RNA/DNA extraction of Frozen Tissue Sections; Preparation and LCM of Paraffin Embedded Tissue Sections; Standard Protocols for Microdissected Tissue Analysis. - [Read Protocols for LCM Preparation and Analysis]

Protocols for Mass Spectrometry Analysis Preparation - http://www.st-andrews.ac.uk/~bmsmspf/protocols.htm#manual

Category: Mass Spectrometry Protocols

Protocols for Mass Spectrometry Analysis Preparation. Preparing gel slices for PMF or MS/MS , mass spec data interpretation, Submitting proteins in solution, Interpretation of LC-MS/MS data. BMS MASS SPECTROMETRY AND PROTEOMICS FACILITY - [Read Protocols for Mass Spectrometry Analysis Preparation]

Protocols for the Maintenance of Human Embryonic Stem Cells in Feeder Free Conditions - http://www.geron.com/PDF/scprotocols.pdf

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

Includes protocols: Mouse Embryonic Fibroblasts (MEF) Preparation; Harvesting MEFs; Cryopreservation of MEFs; Thawing and maintaining MEFs; Irradiating & Plating MEFs; Culture of Human ES cells with Matrigel® and Conditioned Medium; Preparation of Conditioned Medium (CM); Preparation of Matrigel® -coated plates; Passage of human ES cells on Matrigel®; Daily maintenance of feeder-free culture; Freezing Human ES Cells; Thawing Human ES cells; Formation of Embryoid Bodies; - [Read Protocols for the Maintenance of Human Embryonic Stem Cells in Feeder Free Conditions]

Protoplast Preparation for A nidulans Protocol - http://www.aspergillus.org.uk/indexhome.htm?secure/laboratory_protocols/./protoplastRL.html~main

Category: Fungi Protocols: Aspergillus Protocols

Protocol for protoplast preparation for A nidulans. - [Read Protoplast Preparation for A nidulans Protocol]

Purification of Antibodies on an Antigen Column Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4284

Category: Chromatography Protocols: Immunoaffinity Chromatography Protocols

Immunoaffinity purification of antibodies is used to purify antigen-specific antibodies from a preparation of polyclonal antibodies. Such purification is commonly needed in the production of antipeptide antibodies, where it is used to concentrate the desired antibodies and separate them from those raised against carrier proteins. It is also used for the more general purpose of removing unwanted, nonspecific binding activity from polyclonal antibody preparations. - [Read Purification of Antibodies on an Antigen Column Protocol]

Purification of Bovine Lens and Bacterially Expressed Human Vimentin Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4675

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 a method for purifying one type of IF, vimentin, from bovine lens tissue. Purification of human vimentin expressed in Escherichia coli is also described. These methods are useful in the preparation of other IF protein subunits for microinjection studies as well. - [Read Purification of Bovine Lens and Bacterially Expressed Human Vimentin Protocol]

Purification of PCR Products in Preparation for Cloning Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3825

Category: Cloning Protocols

Protocol describes how to use proteinase K to destroy thermostable enzymes and to purify amplified DNA in preparation for cloning. - [Read Purification of PCR Products in Preparation for Cloning Protocol]

Purification of PCR Products in Preparation for Cloning Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3825

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols

This protocol describes how to use proteinase K to destroy thermostable enzymes and to purify amplified DNA in preparation for cloning. - [Read Purification of PCR Products in Preparation for Cloning Protocol]

Quantitative PCR Protocol II - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3842

Category: PCR Protocols: Quantitative PCR Protocols

Quantitative PCR involves co-amplification of two templates: a constant amount of a preparation containing the desired target sequence and serial dilutions of a reference template that is added in known amounts to a series of amplification reactions. The concentration of the target sequence is determined by simple interpolation into a standard curve. - [Read Quantitative PCR Protocol II]

Rapid and Reliable Preparation of Total Cell Lysates from Fission Yeast Protocol - http://www.natureprotocols.com/2006/07/21/rapid_and_reliable_preparation.php

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Protein Purification and Isolation Protocols

Protocol for rapid and reliable preparation of total cell lysates from fission yeast. Method for successful and reproducible extraction of proteins from S. pombe cells. - [Read Rapid and Reliable Preparation of Total Cell Lysates from Fission Yeast Protocol]

Rapid Method for Preparation of Genomic DNA from P. aeruginosa Protocol - http://wheat.pw.usda.gov/~lazo/methods/goldberg/genomic.html

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: Genomic DNA Isolation Protocols

Protocol for rapid method for preparation of genomic DNA from P. aeruginosa. - [Read Rapid Method for Preparation of Genomic DNA from P. aeruginosa Protocol]

Rapid Preparation of DNA from Cells in 96-Well Tissue Culture Dishes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4413

Category: Stem Cell Protocols

This protocol describes a method for rapid preparation of DNA from cells in 96-well tissue culture dishes. - [Read Rapid Preparation of DNA from Cells in 96-Well Tissue Culture Dishes Protocol]

Reactivating Cell Lines and Cell Growth for DNA Preparation - http://humgen.wustl.edu/hdk_lab_manual/hcc/hcc4.html

Category: Cell Biology and Cell Culture: Cell Culture Techniques: Cell Line Preservation Freezing

Cell lines are reactivated and grown to a count of 1 x 108 cells. The cells are pelleted and stored frozen at -80 degrees C prior to DNA extraction. Donis-Keller. - [Read Reactivating Cell Lines and Cell Growth for DNA Preparation]

Real-Time or Kinetic PCR - http://dna-9.int-med.uiowa.edu/realtime.htm

Category: Real Time PCR

Real-Time Chemistry, Instrumentation, Real-Time PCR Quantitation, Quantitative Primer and Probe Design, Thermal Cycling Parameters, Sample Preparation, and Real-time Data Analysis. - [Read Real-Time or Kinetic PCR]

Real-Time RT-PCR: cDNA Synthesis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4114

Category: PCR Protocols: Real-Time PCR Protocols

Protocol uses the Superscript II First-Strand Synthesis system for the generation of cDNA from total RNA. RNA purified using TRIzol reagent (Invitrogen) or the methods described in Preparation of RNA Using Guanidinium Isothiocyanate/Cesium Chloride Ultracentrifugation, Preparation of RNA from Paraffin-Embedded Fixed Tissue. - [Read Real-Time RT-PCR: cDNA Synthesis Protocol]

Real-Time RT-PCR: cDNA Synthesis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4114

Category: PCR Protocols: Reverse Transcriptase RT-PCR Protocols

Removal of Cross-reactive Antibodies from Antiserum: Affinity Chromatography Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3340

Category: Chromatography Protocols: Immunoaffinity Chromatography Protocols

Protocol describes a method for removing antibodies that react with bacterially encoded proteins by passing a crude preparation of immunoglobulins through a column containing immobilized bacterial proteins. - [Read Removal of Cross-reactive Antibodies from Antiserum: Affinity Chromatography Protocol]

Retrieval of DNA Fragments from Pulsed-field Gels following DNA Concentration Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4035

Category: Nucleic Acid Electrophoresis Protocols: Pulsed Field Gel Electrophoresis Protocols

Protocol for retrieval of DNA fragments from pulsed-field gels following DNA concentration. DNA contained in a slice of low-melting-temperature agarose is first concentrated by electrophoresis into a high-percentage agarose gel, and then isolated by treatment with agarase. The resulting DNA preparation is purified by microdialysis. - [Read Retrieval of DNA Fragments from Pulsed-field Gels following DNA Concentration Protocol]

Riboprobe In Vitro Transcription Systems - http://www.promega.com/tbs/tm016/tm016.pdf

Category: Cell Biology and Cell Culture: In Vitro Reconstitution Protocols

In vitro transcription systems includes instructions for use of products P1420, P1430, P1440, P1450 and P1460.Includes information and protocols on RNA Transcription in vitro. Information on DNA Template Preparation;Synthesis of High-Specific-Activity Radio labeled RNA Probes;Determining Percent Incorporation and Probe Specific Activity;Removal of the DNA Template Following Transcription;Removal of unincorporated nucleotides;Synthesis of large amounts of RNA;Capping RNA for in vitro translation. - [Read Riboprobe In Vitro Transcription Systems]

RNA In Situ Hybridization of Dissected Gonads with Digoxygenin-Labeled Probes Protocol - http://www.genetics.wustl.edu/tslab/Protocols/gonad_in_situ.html

Category: C. elegans Protocols

Protocol for RNA in situ hybridization of dissected gonads with digoxygenin-labeled probes. Includes: Preparation of single-stranded probes from cloned cDNA; Dissections and Fixation; Proteinase K digestion; Hybridization; Making a Mount. - [Read RNA In Situ Hybridization of Dissected Gonads with Digoxygenin-Labeled Probes Protocol]

RNA in situ Probe Synthesis Protocol - http://www.fhcrc.org/science/labs/moens/Protocols/probe.html

Category: Cytogenetics Protocols: RNA In Situ Hybridization Protocols

Protocol for RNA in situ probe synthesis. Includes:DNA Template Preparation; Probe Synthesis; Probe Purification using a Sephadex G-50 Spin Column; Probe Hydrolysis. - [Read RNA in situ Probe Synthesis Protocol]

RNA interference protocol - http://www.mann.embl-heidelberg.de/GroupPages/PageLink/activities/iTAP/Experimental/RNAi.html

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]

RNA Preparation from Cultured Cells or Tissue Samples Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/R2.html

Category: Nucleic Acid Purification Protocols: RNA Isolation Protocols: RNA Isolation fromTissues or Cultured Cells Protocols

Protocol for RNA preparation from cultured cells or tissue samples. This protocol has been used to isolate RNA from relatively small tissue samples. The RNA is clean enough for Rnase protection, cDNA synthesis, and RT-PCR analysis. - [Read RNA Preparation from Cultured Cells or Tissue Samples Protocol]

RNA Whole Mount In Situ Hybridization Protocol - http://genetics.med.harvard.edu/~cepko/protocol/ctlab/ish.ct.htm

Category: Cytogenetics Protocols: In Situ Hybridization Tissue Sections Protocols

Protocol for RNA whole mount in situ hybridization. Includes: Embryo preparation; Prehybridization and Hybridization; Post-hybridization washes, blocking, and antibody incubation; Post-antibody washes; Color development. - [Read RNA Whole Mount In Situ Hybridization 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.