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Single Tube Confirmation Yeast PCR Protocol - http://sequence-www.stanford.edu/group/yeast_deletion_project/single_tube_protocol.html

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

Protocol for single tube confirmation yeast PCR. Includes: Clonal purification; Zymolyase treatment; PCR reactions; PCR conditions; Agarose gel electrophoresis. - [Read Single Tube Confirmation Yeast PCR Protocol]

Summary of Methods using Pre-formed Iodixanol Gradients for the Purification of Virsues. - http://www.axis-shield.com/densityhome/optiprep/M17.pdf

Category: Cell Biology and Cell Culture

Summary of the techniques used for the purification of viruses using pre-formed iodixanol gradients. Viruses purified include: Semliki Forest, Retrovirus (from human melanoma cells), Rabies, Polyoma, Papillomavirus, Lassa, Influenza, Bovine viral diarrhea. - [Read Summary of Methods using Pre-formed Iodixanol Gradients for the Purification of Virsues.]

Synthesis of Radiolabeled, Subtracted cDNA Probes Using Oligo(dT) as a Primer Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3869

Category: PCR Protocols: cDNA Synthesis Protocols

Protocol describes the preparation of subtracted cDNA probes by hybridization to an mRNA driver, followed by purification of the single-stranded radiolabeled cDNA by hydroxyapatite chromatography. Before preparing the probe, it is a good idea to have filters (which contain the cDNA library to be screened) ready to hybridize. - [Read Synthesis of Radiolabeled, Subtracted cDNA Probes Using Oligo(dT) as a Primer Protocol]

Tandem Affinity Purification (TAP) Protocol - http://depts.washington.edu/~yeastrc/ms_tap1.htm

Category: Mass Spectrometry Protocols

Tandem Affinity Purification (TAP) Protocol. Modified from Rigaut et al. 1999. Univ.Washington Yeast Resource Center. - [Read Tandem Affinity Purification (TAP) Protocol]

TAP Fusion Protein Purification from Yeast Protocol - http://www.med.unc.edu/~hdohlman/TAP.html

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Immunoprecipitation Protocols

Protocol for TAP fusion protein purification from Yeast. - [Read TAP Fusion Protein Purification from Yeast Protocol]

TEV Purification Protocol - http://openwetware.org/wiki/Keating:Experimental_Protocols:TEV_purification

Category: Viral Protocols

Protocol for TEV purification. - [Read TEV Purification Protocol]

Understanding Protein Trafficking in Plant Cells Through Proteomics - http://www.cepceb.ucr.edu/resources/pdf/pan/ERP-review2_781_2005.pdf

Category: Protein Protocols: Protein Trafficking

Background and methods to study plant transport. Includes new methods to study protein trafficking in plant cells, includes: Identification of protein sorting pathways in non purified samples; Localization of organelle proteins by isotype tagging/isotype-coded affinity tag; Coupling of chemical genomics and proteomics; Top down mass spectrometry; Compartment-specific markers to aid in the purification of organelles. - [Read Understanding Protein Trafficking in Plant Cells Through Proteomics]

UV Absorbance 280 nm Protein Determination - http://wolfson.huji.ac.il/purification/Protocols/OD280.html

Category: Protein Protocols: Protein Quantitation Concentration Protocols

UV Absorbance 280 nm Protein Determination. Simple and quick method to accurately quantitate total protein in purified material or approximately quantitate total protein in crude lysates or partial purified material. Quantitation of the amount of protein in a solution is possible in a simple spectrometer. Absorption of radiation in the near UV by proteins depends on the Tyr and Trp content (and to a very small extent on the amount of Phe and disulfide bonds). Dr. Mario Lebendiker - [Read UV Absorbance 280 nm Protein Determination]

Western Blot Analysis of Endogenous Gene Expression Protocol - http://www.flemingtonlab.com/Protocols/WestBlotAnalyofEndogProts.pdf

Category: Western Blot Protocols

Procedure describes how it denatures most of the modification and degradation proteins immediately giving the most accurate read out of the true levels of protein at the time of harvest. However, in cases where detection is a problem, a limited purification (e.g. isolation of nuclear extract for the detection of transcription factors) might be required to allow analysis. - [Read Western Blot Analysis of Endogenous Gene Expression Protocol]

Working with Bacteriophage P1 and Its Cloning Systems Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4003

Category: Viral Manipulation Protocols: Phage Protocols

Protocol describes methods for recovery and purification of recombinant clones of bacteriophage P1 or PAC DNAs from bacteria. Because of their large size, these DNAs are sensitive to shearing forces and must be handled carefully. This protocol generally yields P1 DNA that works well as a substrate or template in enzymatic reactions. - [Read Working with Bacteriophage P1 and Its Cloning Systems Protocol]

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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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