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(LCM): Preparation and Sectioning of Frozen Tissue Blocks and Purification of RNA from Isolated Cel - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4107

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols: Tissue Preparation for LCM Protocols

LCM isolates specific cells or tissues from samples mounted on microscope slides. The samples are viewed through a thermoplastic film that is attached to a microcentrifuge tube lid. Localized heat, caused by the application of a laser pulse, fuses the membrane to the cells of interest, which can then be harvested for further analysis. RNA and proteins can be purified from the isolated cells, allowing detailed analysis of gene expression. This protocol is divided into three stages. - [Read (LCM): Preparation and Sectioning of Frozen Tissue Blocks and Purification of RNA from Isolated Cel]

96-Well Confirmation Yeast PCR Protocol - http://sequence-www.stanford.edu/group/yeast_deletion_project/96_well_confirmation.html

Category: Yeast Protocols: Yeast Nucleic Acid Protocols: Yeast PCR Protocols

Protocol for 96-well confirmation Yeast PCR. Includes: Clonal purification; Generate a master plate (96-well format); Making a frozen backup stock; Confirmation PCR for one Row; ORF Specific Confirmation PCR --> "A-B" primers (upstream junction); Transfer template DNA to multiwell PCR plate; Prepare and dispense master mix for A-B PCR. - [Read 96-Well Confirmation Yeast PCR Protocol]

A high yield affinity purification method for specific RNA-binding proteins. - http://www.pubmedcentral.gov/pagerender.fcgi?tool=pmcentrez&pageindex=1&artid=330202

Category: RNA Protocols: RNA-binding Protein Purification

A high yield affinity purification method for specific RNA-binding proteins: isolation of the iron regulatory factor from human placenta. Neupert 1990. - [Read A high yield affinity purification method for specific RNA-binding proteins.]

A high-capacity RNA affinity column for the purification of human IRP1 and IRP2 overexpressed - http://www.rnajournal.org/cgi/reprint/9/3/364.pdf

Category: Chromatography Protocols: DNA RNA Affinity Chromatography

PDF. A high-capacity RNA affinity column for the purification of human IRP1 and IRP2 overexpressed in Pichia pastoris. CHARLES R. ALLERSON, ALAN MARTINEZ, EMINE YIKILMAZ and TRACEY A. ROUAULT. RNA Journal. - [Read A high-capacity RNA affinity column for the purification of human IRP1 and IRP2 overexpressed]

A Method for Isolation of Chloroplast DNA and Mitochondrial DNA from Sunflower - http://pubs.nrc-cnrc.gc.ca/ispmb/ispmb16/16183-2.pdf

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

The protocol includes: organelle isolation, deoxyribonuclease treatment, lysis, deproteinisation and a final DNA purification with sodium dodecyl sulphate and potassium acetate. The organelle DNA yield is 5–10 micrograms per gram of tissue and the DNA is fully restrictable. The technique is inexpensive and appropriate for the isolation of multiple samples of organelle DNA from a small amount of tissue. - [Read A Method for Isolation of Chloroplast DNA and Mitochondrial DNA from Sunflower]

A Method for Micro-Scale Isolation and Purification of Gangliosides - http://www.glycotech.com/protocols/Proto1.html

Category: Molecular Biology Protocols: Carbohydrate Protocols

Solvent partition protocol allows the isolation of gangliosides from small samples and from samples where ganglioside concentrations are low, especially relative to the concentration of potentially contaminating proteins and other large molecular weight species. Stephan Ladisch Director, Center for Cancer and Transplant Biology, Children's National Medical Center, Washington, D.C. - [Read A Method for Micro-Scale Isolation and Purification of Gangliosides]

A Protocol for AAV Vector Production and Purification - http://www.brc.riken.jp/lab/dna/rvd/SOP/AAV/AAVProtocol.pdf

Category: Viral Culture and Isolation Protocols: Viral Isolation Protocols

Protocol describes typical methods that are used to propagate and purify AAV vectors for experiments both in vitro and in vivo. Includes: Principles of the Triple Plasmid Transfection System; Plasmids; Transfection and Extraction of Virus; Purification of the AAV vector. - [Read A Protocol for AAV Vector Production and Purification]

AAV Vector Production and Purification Protocol - http://www.brc.riken.jp/lab/dna/rvd/SOP/AAV/AAVProtocol.pdf

Category: Cloning Protocols: Vector Protocols

Protocol is used to propagate and purify AAV vectors for experiments both in vitro and in vivo. Includes: Principles of the Triple Plasmid Transfection System; Plasmids; Transfection and Extraction of Virus; Purification of the AAV vector. - [Read AAV Vector Production and Purification Protocol]

ABI Prism 310 Cycle Sequencing Protocol - http://vosshall.rockefeller.edu/protocols/ABICYCLESEQ.pdf

Category: DNA Protocols: DNA Sequencing Protocols

Protocol for ABI prism 310 cycle sequencing. Includes: Sequencing Reaction; Column Purification; Sample Loading Preparation. - [Read ABI Prism 310 Cycle Sequencing Protocol]

Affinity Antibody Purification of Protein A/G Chromatography - http://www.upstate.com/misc/protocol_detail.q.prot.e.antibody_purification.a.name.e.Antibody_Purification

Category: Chromatography Protocols: Immunoaffinity Chromatography Protocols

Antibody Purification (Antiserum or Ascites by Protein A/G Chromatography). Species and Type of Antibody Agarose Rabbit IgG Protein A or Protein G Mouse IgG1 Protein G Mouse IgG2 Protein A or Protein G Mouse IgG3 Protein G Sheep IgG Protein G but binds weakly Rat IgG Protein G but binds weakly Guinea Pig IgG Protein A Dog IgG Protein A Goat IgG Protein G Pig IgG Protein A Hamster IgG Protein G. By Millipore. - [Read Affinity Antibody Purification of Protein A/G Chromatography]

Affinity Chromotography Magnetic Bead Purification of RNA-binding proteins from Rat Brain. - http://147.52.72.117/IJMM/2003/volume11/number4/509.pdf

Category: RNA Protocols: RNA-binding Protein Purification

Affinity Chromotography Magnetic Bead Purification of RNA-binding proteins from Rat Brain. Scaturro et al. 2003. - [Read Affinity Chromotography Magnetic Bead Purification of RNA-binding proteins from Rat Brain.]

Affinity Purification of Antibodies - http://cmgm.stanford.edu/~vesicle/protocols/affinity.html

Category: Protein Protocols: Antibody Purification Protocols

Affinity Purification of Antibodies. Vesicle Trafficking. - [Read Affinity Purification of Antibodies]

Affinity purification of antibodies from crude serum - http://wheat.pw.usda.gov/~lazo/methods/au/sect32.html

Category: Protein Protocols: Antibody Purification Protocols

Affinity purification of antibodies from crude serum. David Bowtell Lab. - [Read Affinity purification of antibodies from crude serum]

Affinity purification of antibodies Protocol - http://www.xenbase.org/xmmr/Marker_pages/methods_pages/affinity_col.html

Category: Protein Protocols: Antibody Purification Protocols

Affinity purification of antibodies. Protocol by Peter Vize, XMMR - [Read Affinity purification of antibodies Protocol]

Affinity Purification of Interacting Proteins from Cell Lysates Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4593

Category: Chromatography Protocols: Affinity Chromatography Protocols

Recombinant protein or a chemically synthesized bioactive fragment is immobilized on resin and used as a probe to capture interacting proteins directly from a cell extract. Affinity-purified proteins are fractionated by gel electrophoresis and visualized by Coomassie staining. Proteins that interact specifically are identified by comparing this gel profile to one obtained from cell lysates passed over a control resin lacking the immobilized probe protein. - [Read Affinity Purification of Interacting Proteins from Cell Lysates Protocol]

Alkaline Lysis Mini-Prep Protocol - http://preuss.bsd.uchicago.edu/protocols/Alkaline.html

Category: Microbiology: Mini-Prep Plasmid Purification

Harvesting and Lysis of Bacteria. Mini-prep plasmid purification using Solution I, Solution II and Solution III. The Preuss Lab. Univ. Chicago. - [Read Alkaline Lysis Mini-Prep Protocol]

Antibody Purification - http://axon.med.harvard.edu/~cepko/protocol/mike/A1.html

Category: Protein Protocols: Antibody Purification Protocols

This protocol includes an ammonium sulfate cut, affigel blue chromatography and affinity chromatography. Mike A. Dyer Lab Harvard. - [Read Antibody Purification]

Antibody Purification - Antiserum or Ascites by Protein A/G Chromatography - http://www.upstate.com/misc/protocol_detail.q.prot.e.antibody_purification.a.name.e.Antibody_Purification

Category: Protein Protocols: Antibody Purification Protocols

Antibody Purification (Antiserum or Ascites by Protein A/G Chromatography). Buffer Preparation, Preparation of a Protein A Agarose or Protein G Agarose Affinity Column, Pouring the Protein A/G Affinity Column, Preparation of Antiserum or Ascites for Affinity Chromatography, Affinity Chromatography Using Protein A/G Agarose. - [Read Antibody Purification - Antiserum or Ascites by Protein A/G Chromatography]

ANTIBODY PURIFICATION by affinity chromatography - http://mullinslab.ucsf.edu/protocols/html/ANTIBODY_PURIFICATION_by_affini.htm

Category: Chromatography Protocols: Immunoaffinity Chromatography Protocols

ANTIBODY PURIFICATION by affinity chromatography. By Beth, Mullins Lab UCSF. To affinity purify antibodies, generate lots of E. coli lysate that contains your antigen. If the protein can stand freeze thawing, then go ahead and purify the protein from e. coli lysate and keep it frozen until you need to couple it to a CH-sepharose column. - [Read ANTIBODY PURIFICATION by affinity chromatography]

ANTIBODY PURIFICATION ON THE PROTEIN G COLUMN - http://www.its.caltech.edu/%7Ebjorker/Antibody_purif%20_Protein_G%20.pdf

Category: Protein Protocols: Antibody Purification Protocols

ANTIBODY PURIFICATION ON THE PROTEIN G COLUMN. Jorker Lab, Caltech. - [Read ANTIBODY PURIFICATION ON THE PROTEIN G COLUMN]

Antibody Purification Protocol - http://iacf.bsd.uchicago.edu/Monoclonal_Antibody_Facility/AntibodyPurificationProtocol.htm

Category: Protein Protocols: Antibody Purification Protocols

Antibody Purification Protocol. Monoclonal Antibody Facility - [Read Antibody Purification Protocol]

Antibody Purification using Ammonium Sulphate Precipitation - http://www.icp.ucl.ac.be/mexp/vipg_pur.html

Category: Protein Protocols: Antibody Purification Protocols

Antibody Purification using Ammonium Sulphate Precipitation. Viral Immunity and Pathogenesis Group. - [Read Antibody Purification using Ammonium Sulphate Precipitation]

Antigen Design and Sera Purification - http://www.sigmaaldrich.com/Brands/Sigma_Genosys/Custom_Antisera/Key_Resources/Antigen_Design.html

Category: Immunology: Antigen

Antigen Design and Sera Purification. Custom antisera. Sigma Aldrich. Peptide Selection and Design, Coupling Strategy # Selecting the Protein Carrier, Multiple Antigenic Peptides (MAPs), Choice of Host, Adjuvant, Immunization, & Sera Collection, Antisera Purification, Ammonium Sulfate Precipitation, Protein A/G, Immunoaffinity Purification. - [Read Antigen Design and Sera Purification]

BACs Bacterial Artificial Chromosomes - http://openwetware.org/wiki/Bacterial_artificial_chromosomes

Category: BACs Bacterial Artificial Chromosome Protocols

Information for BACs, Bacterial Artificial Chromosomes. Includes: Existing vectors; pBAC108L; pBeloBAC11; pBACe3.6; pFW11 and F' factor; pCC1BAC; pSMART VC Vectors; pIndigoBAC-5; Cosmids; pWEB-TNCTM; SuperCos 1; pFos1; PAC vectors; BAC purification; Putting the BAC into E. coli. - [Read BACs Bacterial Artificial Chromosomes]

C. elegans RNAi Protocol - http://www.zoology.ubc.ca/~alorch/rnai-protocol.htm

Category: C. elegans Protocols: C. elegans Genetics Protocols

Protocol for C. elegans RNAi. Includes: Transformation of competent cells; Blunt-end ligation; Preparation of competent cells; Dephosphorylation of linear plasmid DNA; Restriction Digest: EcoRV; Insert amplification from gDNA; Gel purification: QiaQuick gel purification kit; Mini-prep; Transformant Screening; Bacterial preparation and induction; Preparation of worms for RNAi feeding. - [Read C. elegans RNAi Protocol]

Articles (1-4 of 4)

Genomic DNA Labeling of Microarrays Protocol

DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.

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.

DNA Fragment Purification from Polyacrylamide Gels Protocol

A simple protocol for the purification of DNA fragments from Polyacrylamide Gels.

Electrotransformation of BMH 81-17mut S for Isolating Site-Directed dsDNA Mutants

This protocol describes the electroporation of the BMH 81-17 mut S strain that is recommended for tranformation of the site directed mutagenesis of dsDNA (See Protocol on Site-Directed Mutagenesis on Double Stranded DNA). BMH 81-17 mut S are a mismatch repair defective (mut S) Escherichia coli strain. The probability that the two mutations will cosegregate during the first round of DNA replication is increased in this strain.

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