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2-D Polyacrylamide Gel Electrophoresis Protocol - http://cgap-mf.nih.gov/Protocols/DNARNAProteomicAnalysis/Proteomics/2DPage.html

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

Protocol for 2-D polyacrylamide gel electrophoresis. Includes: 50 ml IEF Lysis Buffer; 10X Electrophoresis Running Buffer (10 L); 30% Acrylamide Stock (1 liter); Separating Acrylamide Gel; 50 ml Equilibration Buffer I; 50 ml Equilibration Buffer II; Transfer Buffer; Sample Preparation; Tissue Processing; Reswelling; Prepare Gradient Acrylamide Gel (9-18%); Transfer and Sequencing of Proteins. - [Read 2-D Polyacrylamide Gel Electrophoresis Protocol]

Culturing of Insect Cells and Production of Baculovirus Expressed Recombinant Proteins in 2.8 Liter - http://www.hyclone.com/pdf/procedure_insect_fernbach.pdf

Category: Cell Culture Protocols: Insect Cell Culture Protocols

The use of Ferbach flasks is an ideal method to amplify the cells and virus inoculum for the bioreactor. The method is also very practical for producing sufficient amounts of various proteins for feasibility studies using HyQ© SFX-Insect™ medium. The large surface area in this vessel enables sufficient aeration and, therefore, the cells, when they reach high density, are not oxygen depleted. - [Read Culturing of Insect Cells and Production of Baculovirus Expressed Recombinant Proteins in 2.8 Liter]

Preparation of KC Nuclear Extract for In Vitro Splicing - http://www.bio.com/protocolstools/protocol.jhtml%3Bjsessionid%3D34E3CBZRBQMFZR3FQLMCFEWHUWBNSIV0?id=p521

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

Protocol for preparation of KC nuclear extract for in vitro splicing. Protocol makes 3.4 ml of extract for every 4 liter of cells (depending on initial cell concentration). Protocol includes: Procedure, Solutions, BioReagents and Chemicals and protocol hints. - [Read Preparation of KC Nuclear Extract for In Vitro Splicing]

Preparing Vacuolar Membranes And Other Membrane Fractions from Neurospora Crassa Protocol - http://www.fgsc.net/neurosporaprotocols/VM%20vacuole%20procedure.pdf

Category: Fungi Protocols: Neurospora Protocols

Protocol for Preparing Vacuolar Membranes And Other Membrane Fractions from Neurospora Crassa (4 liter prep, easily scaled up to 10, 12, 24, or 30 l). - [Read Preparing Vacuolar Membranes And Other Membrane Fractions from Neurospora Crassa Protocol]

Staining Chromosomes (G-Banding) Protocol - http://humgen.wustl.edu/hdk_lab_manual/cam/cam4.html

Category: Cytogenetics Protocols

Protocol for staining chromosomes (G-Banding). Includes: Trypsin (1:250) USP Grade; Gurr Buffer Solution: (1 liter); Working Salt Solution: (1 liter); Leishman Stain Solution. - [Read Staining Chromosomes (G-Banding) Protocol]

Wet-lab protocols - http://eagle.mmid.med.ualberta.ca/private/labnotes/wetlab/index.html

Category: Microbiology: Bacterial E.Coli Culture Media & Plates

LB plates, LB medium (for 1 liter), Agarose gels. Bart.Hazes Univ Alberta. - [Read Wet-lab protocols]

Yeast Plates Protocols - http://www.fhcrc.org/science/labs/hahn/methods/genetic_meth/yeast_plates.html

Category: Yeast Cell Culture Protocols: Yeast Media, Solutions and Stocks Protocols

Yeast plates protocols. Includes: Glucose Complete Plates (1 liter); YPD Plates (1 liter); 5-FOA plates (0.5 liter). - [Read Yeast Plates Protocols]

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Lambda Phage Protocol Large DNA Preparation

Single Stranded Plasmid DNA Isolation Protocol

A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.

Antibodies Phage Expression Protocol Using 96-well Microtiter Plates

A Phage display expression protocol for antibody expression in 96-well microtiter plates.

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.

Coupling Cysteine Containing Peptides to Carrier Protein for Immunization and Polyclonal Antibody

This protocol is used in the production of polyclonal antibodies that recognize a peptide sequence of interest.

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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