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LB Plates Pouring and Preparation - http://www.bbri.org/faculty/smith/LB_Plates.html

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

LB Plates Pouring and Preparation. antibiotics addition. Janet Smith, Boston Biomedical Research Institute - [Read LB Plates Pouring and Preparation]

Liquid Culture of Worms Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=22

Category: C. elegans Protocols: C. elegans Cell Culture Protocols

Protocol for liquid culture of worms. Includes: superbroth; S- basal; worm plates; GROWING THE BACTERIA (WORM FOOD); GROWING THE WORMS; AFTER THE CULTURE HAS GROWN; PREPARING EGGS TO START SYNCHRONIZED LIQUID CULTURES. - [Read Liquid Culture of Worms Protocol]

Luciferse Assay Protocol for 12 Well Plates. Word Doc. - http://www.med.umich.edu/intmed/endocrinology/hammerlab/Protocols/Luciferase%20Assay%20Protocol.doc

Category: Luciferase and Dual Luciferase Assays

Luciferse Assay Protocol for 12 Well Plates. Word Doc. Hammer Lab. - [Read Luciferse Assay Protocol for 12 Well Plates. Word Doc.]

MAKING MICROARRAY PRINTING PLATES (IN DMSO) - http://pga.tigr.org/sop/M003_1a.pdf

Category: DNA Microarray Protocols

This protocol describes the method for making microarray printing plates in a 96 well format. (The same procedure applies to a 384 well format as well.) Hasseman. TIGR Microarray Protocols - [Read MAKING MICROARRAY PRINTING PLATES (IN DMSO)]

Mammalian Cell Culture and Preparation of Cells for RNAi in 24-Well Plates Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4341

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

This protocol introduces a method for maintaining mammalian cell culture suitable for treatment with siRNA. Transfection of cultured cells with siRNAs (see Transfection of siRNA Duplexes into Mammalian Cells) and downstream analysis of the knockdown cells. - [Read Mammalian Cell Culture and Preparation of Cells for RNAi in 24-Well Plates Protocol]

Mammalian Cell Culture and Preparation of Cells for RNAi in 24-Well Plates Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4341

Category: RNAi Technology Protocols: RNAi Mammalian Cells Protocols

Protocol introduces a method for maintaining mammalian cell culture suitable for treatment with siRNA. Transfection of cultured cells with siRNAs and downstream analysis of the knockdown cells. - [Read Mammalian Cell Culture and Preparation of Cells for RNAi in 24-Well Plates Protocol]

Measurement of CAT in Extracts of Mammalian Cells Using Thin-layer Chromatography - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3951

Category: Reporter Gene Assays: Cat Assays Protocols

In this protocol, extracts prepared from cells transfected with a chloramphenicol acetyltransferase (CAT) reporter plasmid are incubated with radiolabeled chloramphenicol. The acetylated products generated by the action of CAT are separated from the unmodified drug by thin-layer chromatography and quantitated by scraping the spots from the thin-layer plates and counting them by scintillation spectroscopy. - [Read Measurement of CAT in Extracts of Mammalian Cells Using Thin-layer Chromatography]

Mini-Chamber for Regulating Gaseous Environment During Culture - http://www.animal.ufl.edu/hansen/protocols/minigaschamber.htm

Category: Developmental Biology: Embryology Protocols

A simple and inexpensive chamber for regulating gaseous environment of small culture plates, such as those used for culture of preimplantation embryos, can be constructed using disposable media-filtration devises such as Corning’s 115-ml system. The following is a description of how to make such a device. - [Read Mini-Chamber for Regulating Gaseous Environment During Culture]

Passage Procedure for RAW 264.7 Cells - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000159.pdf?pid=PP00000159

Category: Cell Culture Protocols: Cell Lines Protocols

RAW 264.7 cells are a macrophage-like, Abelson leukemia virus transformed cell line derived from BALB/c mice. For routine maintenance in culture (passage), cells are seeded at a confluence of approximately 10% (1 x 106 and 3 x 106 cells in 100-mm and 150-mm plates, respectively) and grown to a confluence of approximately 80%. This procedure requires the cells to be split every two days. - [Read Passage Procedure for RAW 264.7 Cells]

Pouring Plates - http://rothlab.ucdavis.edu/protocols/M10.html

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

Pouring Plates. Bacterial culture LB. John Roth, U.C Davis. - [Read Pouring Plates]

Preparation of Broth and Plates - http://www.people.virginia.edu/~gwl6s/protocols/recipes.html

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

Terrific Broth, LB plates, NZY Plates, LB Broth. Laurie Lab, Univ. Virginia Health Science Center. - [Read Preparation of Broth and Plates]

Preparation of DNA from Cells in 24-Well Tissue Culture Plates Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4414

Category: Stem Cell Protocols

This protocol describes a method of DNA preparation from ES cells in 24-well plates. - [Read Preparation of DNA from Cells in 24-Well Tissue Culture Plates Protocol]

Preparation of Mouse Embryonic Fibroblast (MEF) Feeder Plates Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4483

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

MEF feeders are prepared weekly to provide a substrate for undifferentiated embryonic stem (ES) cells. Primary MEF cells are thawed, established in culture, treated with mitomycin C to halt their proliferation so they cannot overgrow the ES cultures, and then replated onto dishes convenient for ES cell culture. This protocol can also be used to prepare feeder cells from STO fibroblast cell lines. - [Read Preparation of Mouse Embryonic Fibroblast (MEF) Feeder Plates Protocol]

Preparation of Seeded NGM Plates For Worm Food Protocol - http://www.bio.com/protocolstools/protocol.jhtml?id=p1558

Category: C. elegans Protocols: C. elegans Cell Culture Protocols

Protocol describes how to seed plates with E. coli OP50 for C. elegans cultures. Includes: Bacterial Stock Plate Preparation; Bacterial Broth Preparation; Seeding Bacteria Plates for Culturing Worms. - [Read Preparation of Seeded NGM Plates For Worm Food Protocol]

Preparing the Aggregation Plate Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4420

Category: Mouse Protocols: Assembling Chimeras Mouse Protocols

Protocol describes the preparation of aggregation plates, which will then be used for the production of aggregation chimeras. - [Read Preparing the Aggregation Plate Protocol]

Principles and Applications of Liquid Scintillation Counting PDF - http://www.ehs.psu.edu/radprot/LSC_Theory2.pdf

Category: Radioactivity: Liquid Scintillation Counting

Excellent guide for Liquid Scintillation Counting. Includes protocols and methods for counting gel slices, SPECIAL SAMPLE PREPARATION PROTOCOLS: TLC Plates, protocol for Counting Samples on Cellulose-ester, Filters (MilliporeTM filters), Counting Tissue radioacitivity, Counting 14CO2, Samples in Polyacrylamide Gels. National Diagnostics. National Diagnostics Laboratory Staff. - [Read Principles and Applications of Liquid Scintillation Counting PDF]

Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4484

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the culture of embryonic stem (ES) cells using mitotically inactivating primary mouse embryonic fibroblast (MEF) cells as a feeder layer (preparation described in Preparation of Mouse Embryonic Fibroblast (MEF) Feeder Plates). The ES culture medium is supplemented with recombinant leukemia inhibitory factor (LIF) to help maintain the cells as pluripotent stem cells. This protocol has been optimized for the ES-D3 cell line. - [Read Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol]

Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4484

Category: Stem Cell Protocols: Mouse Embryonic Fibroblasts Protocols

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]

RNA Interference Screening (RNAi) in Cultured Cells - http://flyrnai.org/RNAi_FAQ_rnai_on_cc.html

Category: siRNA and RNAi Protocols

The protocol listed is Drosophila cells in 6 well plates and our pre-aliquoted 384 well plates. RNAi experiments may be done in other size plates, just scale up or down. Drosophila RNAi Screening Center - [Read RNA Interference Screening (RNAi) in Cultured Cells]

Screening Bacterial Colonies by Hybridization: Intermediate Numbers Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3946

Category: Bacterial Protocols

Bacterial colonies growing on agar plates are transferred en masse to nitrocellulose filters. The spatial arrangement of colonies on the plates is preserved on the filters. After transfer, the filters are processed for hybridization to an appropriate radiolabeled probe while the original (master) plate is incubated for a few hours to allow the bacterial colonies to regrow in their original positions. - [Read Screening Bacterial Colonies by Hybridization: Intermediate Numbers Protocol]

Screening Bacterial Colonies by Hybridization: Small Numbers Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3935

Category: Bacterial Protocols

Protocol used to screen a small number of bacterial colonies (<200) that are dispersed over several agar plates and are to be screened by hybridization to the same radiolabeled probe. The colonies are gridded onto a master plate and onto a nitrocellulose or nylon filter laid on the surface of a second agar plate. - [Read Screening Bacterial Colonies by Hybridization: Small Numbers Protocol]

Screening Expression Libraries Constructed in Bacteriophage Lambda Vectors Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3765

Category: DNA Protocols: cDNA Library Protocols: cDNA Library Construction Protocols

An expression library constructed in a bacteriophage {lambda} vector is plated on an appropriate E. coli strain in the absence of isopropylthio-ß-D-galactoside (IPTG). After 2-4 hours, the plates are moved to 37°C (to stabilize any fusion proteins that are temperature sensitive), and filters impregnated with IPTG are laid on top of the developing plaques. - [Read Screening Expression Libraries Constructed in Bacteriophage Lambda Vectors Protocol]

Screening Expression Libraries Constructed in Plasmid Vectors Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4079

Category: DNA Protocols: cDNA Library Protocols

A cDNA library constructed in a plasmid expression vector of the pUC, pUR, or pEX series is plated on agar medium and then replicated onto filters, which are transferred to plates containing IPTG. After 2-4 hours of induction, the colonies are lysed with chloroform and then screened with appropriate antibodies. - [Read Screening Expression Libraries Constructed in Plasmid Vectors Protocol]

SNP Genotyping System - http://www.promega.com/tbs/tm053/tm053.pdf

Category: Genetics and Genomics: Genotyping Protocols

SNP Genotyping System. Includes: Protocol for Manual READIT® Assay in Multiwell Plates; Preparation of L/L Reagent; Preparation of Heating Block; CIAP/Exo Treatment; Preparation of Interrogation Probe Solutions; Template Denaturation; Sample Interrogation; Protocol for Manual Plate Reading Luminometers; Protocol for Injecting Plate Reading Luminometers; Protocol for Automated READIT® Assay with the Biomek® 2000 Instrument. - [Read SNP Genotyping System]

Articles (1-5 of 5)

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.

Picking Transfected ES Cell Clones Protocol

This protocol a protocol on how to generate transfected embryonic stem (ES) cell clones. The previous protocol in this series is the Protocol for Electroporation of ES cells. The next protocol in the series is the Protocol on Disaggregation, Expansion, and Freezing of Transfected ES Clones.

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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