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Isolation of Human NK Subsets Protocol - http://www.natureprotocols.com/2006/08/07/isolation_of_human_nk_subsets.php

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

Methodological details about sorting and purification of peripheral blood and decidual human NK subsets. - [Read Isolation of Human NK Subsets Protocol]

Isolation of Plasmid DNA from Yeast Cells: A Ten-Minute Preparation - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4150

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

Protocol describes the purification release of plasmid DNA from yeast cells. The resulting plasmid DNA can be used to transform Escherichia coli or yeast. - [Read Isolation of Plasmid DNA from Yeast Cells: A Ten-Minute Preparation]

Isolation of Primary Epithelial Cells - http://www.lbl.gov/lifesciences/BissellLab/labprotocols/primarycellprep.htm

Category: Cell Biology and Cell Culture: Epithelial Cell Culture

Isolation of Primary Epithelial Cells. Digestion: In Collagenase Medium, Purification and epithelial cell culture methods. Bissell Lab. - [Read Isolation of Primary Epithelial Cells]

Isolation of Yeast Genomic DNA for Southern Blot Analysis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4149

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

Protocol (modified from Hoffman and Winston 1987) describes the purification of genomic DNA from yeast cells. - [Read Isolation of Yeast Genomic DNA for Southern Blot Analysis Protocol]

Lowry â€“ Protein Determination - http://wolfson.huji.ac.il/purification/Protocols/loury.html

Category: Protein Protocols: Lowry Protein Assay

Lowry â€“ Protein Determination. Quantitation of total protein using the lowry protein assay. Dr. Mario Lebendiker. The Wolfson Centre for Applied Structural Biology, The Hebrew University of Jerusalem. Method materials and notes for the Lowry assay. - [Read Lowry â€“ Protein Determination]

M5 Velocity (rate zonal) gradients for purification and assembly analysis of viruses. - http://www.axis-shield.com/densityhome/optiprep/M05.pdf

Category: Cell Biology and Cell Culture

Purification protocols of the viruses: HIV-1, Lassa virus, oncornavirus and other retroviruses. Protocol uses an iodixanol gradient in a sedimentation velocity mode to purifyHIV-1 virions without affecting the infectivity of the virus. In rate-zonal iodixanol gradients the HIV-1 was effectively separated both from Vif and from the microvesicles. - [Read M5 Velocity (rate zonal) gradients for purification and assembly analysis of viruses.]

Magnetic techniques for the isolation and purification of proteins and peptides - http://www.biomagres.com/content/2/1/7

Category: RNA Protocols: RNA-binding Protein Purification

Magnetic techniques for the isolation and purification of proteins and peptides. Safarik et al. 2004 - [Read Magnetic techniques for the isolation and purification of proteins and peptides]

MICROARRAY PCR, PURIFICATION, AND STORAGE - http://pga.tigr.org/sop/M002_1a.pdf

Category: DNA Microarray Protocols

This protocol describes PCR amplification of eukaryotic cDNA plasmid inserts, gel electrophoresis, purification, and storage of PCR products. Hasseman. TIGR Microarray Protocols. - [Read MICROARRAY PCR, PURIFICATION, AND STORAGE]

Microsome Preparation from Yeast Cells Protocol - http://www.bio.com/protocolstools/protocol.jhtml?id=p1676

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

This protocol describes the isolation of microsomal Endoplasmic Reticulum (ER) membranes from yeast cells. Includes: Spheroplast Formation and Microsome Purification. - [Read Microsome Preparation from Yeast Cells Protocol]

Microsome Purification from Rat Liver Protocol - http://www.bio.com/protocolstools/protocol.jhtml?id=p2051

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

This protocol gives instructions for preparing microsomes from a rat liver. - [Read Microsome Purification from Rat Liver Protocol]

Mini-Prep Protocol for Purification of Plasmid DNA from Bacteria - http://people.morehead-st.edu/fs/d.peyton/protocols.html

Category: Microbiology: Mini-Prep Plasmid Purification

Using Solutions A for Lysis, Solutions B for Neutralization Solution, and Solution C for washing. Dr.David Peyton. - [Read Mini-Prep Protocol for Purification of Plasmid DNA from Bacteria]

mRNA Purification - http://www.fhcrc.org/science/labs/breeden/Methods/mRNA_purification.html

Category: RNA Protocols: mRNA Extraction Protocols

mRNA Purification using oligo dT cellulose. Breeden Lab. - [Read mRNA Purification]

Nick-Translation Labeling of ds DNA with Nick Translation Mixes for In Situ Probes Protocol - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_49-51.pdf

Category: Nucleic Acid Modification Protocols: Nucleic Acid Labeling Protocols

Protocol for nick-translation labeling of ds DNA with nick translation mixes for in situ probes. Includes: Labeling reaction with DIG-dUTP or Biotin-dUTP; Labeling reaction with Fluorescein-dUTP, or Tetramethylrhodamine-dUTP; Purification of labeled probe. - [Read Nick-Translation Labeling of ds DNA with Nick Translation Mixes for In Situ Probes Protocol]

Nonradioactive Cycle Sequencing of PCR-Amplified DNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4095

Category: DNA Protocols: DNA Sequencing Protocols: Big Dye Protocols

Protocol describes the purification, quantification, andsubsequent sequencing of amplified DNA fragments using PCR.Excess nucleotides are removed from the initial PCR productsusing spun columns, and the products are quantified using fluorometry. - [Read Nonradioactive Cycle Sequencing of PCR-Amplified DNA Protocol]

Oligonucleotide Purification Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/O1.html

Category: Nucleic Acid Purification Protocols

Protocol for oligonucleotide purification. - [Read Oligonucleotide Purification Protocol]

One-Step Isolation of Plant DNA Suitable for PCR Amplification - http://pubs.nrc-cnrc.gc.ca/ispmb/ispmb19/r01-054.pdf

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

One step extraction for isolation of plant DNA. DNA suitable for amplification by PCR can be produced from leaf material smaller than 0.3 mm2 in less than 20 min & no tube changes. Method was tested on several plant species. Method was found to extract DNA that could be amplified without any further purification or treatment. The isolated DNA was amplified using a universal chloroplast primer set. The method was validated by comparing size of PCR products generated using standard DNA isolation. - [Read One-Step Isolation of Plant DNA Suitable for PCR Amplification]

Overview of Affinity Purification in Combination with Mass Spectrometry Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/30/pdb.top3

Category: Chromatography Protocols: Affinity Chromatography Protocols

Protein complexes can be isolated by several different approaches. For example, a protein can be tagged with an epitope such as Flag or TAP and then overexpressed in a target cell, allowing the interacting proteins to be purified. Similarly, epitope tags can be homologously recombined into the endogenous locus ("knocked-in"), allowing protein complexes containing the tagged proteins to be isolated at their natural expression level. - [Read Overview of Affinity Purification in Combination with Mass Spectrometry Protocol]

Phycoerythrin Conjugation Protocol - http://pingu.salk.edu/flow/protocols/pelabel.html

Category: Immunology

Protocol for phycoerythrin conjugation. Includes: Preparation of PE; SPDP modification of PE; SMCC modification of antibody; DTT treatment of SPDP-PE; Purification of reactants; Conjugation; Stop reaction; Concentrate product; Separate conjugate. - [Read Phycoerythrin Conjugation Protocol]

Plant Cell Purification of Intact Plant Protoplasts - http://www.axis-shield.com/densityhome/optiprep/C18.pdf

Category: Cell Biology and Cell Culture

Cell fractionation protocol to yield intact plant protoplasts. Technique used to purify protoplasts from the grass Glyceria fluitans. Protocol includes: Determination of leaf osmolality; Sterilization. - [Read Plant Cell Purification of Intact Plant Protoplasts]

Polyclonal Antibody Production Protocol - http://research.cm.utexas.edu/bkitto/Kittolabpage/Protocols/Immunology/PAb.html

Category: Antibody Protocols: Antibody Production Protocols

Protocol for polyclonal antibody production. Very useful for rapid and simple generation of antibodies for western blots, ELISA assays, and immunoprecipitation. Includes: Rabbit Immunization; Initial Preparation; Pre-bleed; Antigen Injection; Monitoring of Titer; Purification of Antibodies. - [Read Polyclonal Antibody Production 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 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 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 cell lysates from yeast by enzymatic digestion of the cell wall - http://www.embl.de/ExternalInfo/protein_unit/draft_frames/protocol_database/Purification/ec_yeast_enzyme.html

Category: Protein Protocols: Protein Extraction From Cells

Preparation of cell lysates from yeast by enzymatic digestion of the cell wall. - [Read Preparation of cell lysates from yeast by enzymatic digestion of the cell wall]

Preparation of cell lysates from yeast using a French Press - http://www.embl.de/ExternalInfo/protein_unit/draft_frames/protocol_database/Purification/ec_yeast_press.html

Category: Protein Protocols: Protein Extraction From Cells

Preparation of cell lysates from yeast using a French Press. he Protein Expression and Purification Facility, EMBL - [Read Preparation of cell lysates from yeast using a French Press]

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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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