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Antigen Retrieval for Immunohistochemistry with Paraffin-Embedded Tissues Protocol - http://www.animal.ufl.edu/hansen/Protocols/Immunohistochemistry_paraffin.htm

Category: Immunohistochemistry Protocols: Antigen Retrieval Protocols

Protocol used to for immunohistochemistry on paraffin-embedded sections. Based on use of microwave energy to effect antigen retrieval. The immunohistochemistry procedure, is for use of Biomeda's HistoScan kit based on a streptavidin-peroxidase/biotinylated second antibody detection system with 3-amino, 9-ethylcarbazole (AEC) as chromogen. Undoubtedly, other kits or home-made reagents will also work . - [Read Antigen Retrieval for Immunohistochemistry with Paraffin-Embedded Tissues Protocol]

Assay for Phage Containing the Beta-galactosidase Gene - http://humgen.wustl.edu/hdk_lab_manual/lambda/lambda7.html

Category: Reporter Gene Assays: B-Galactosidase Assay Protocols

This assay is used when working with phage vectors carrying the beta-gal gene. If the cloning event disrupts a normally functional copy of the gene in the vector the resulting plaques would appear clear in the assay. If the phages contain a functional beta-gal gene they will form blue rings around their plaques. Any strain which is not an overproducer of beta-gal will work as indicator host bacteria; a single chromosomal copy of the gene is not a problem. - [Read Assay for Phage Containing the Beta-galactosidase Gene]

Assay for Phage Containing the Beta-galactosidase Gene Protocol - http://humgen.wustl.edu/hdk_lab_manual/lambda/lambda7.html

Category: Yeast Protocols: Yeast Genetics Protocols: B-Galactosidase Assay Protocols

This assay is used when working with phage vectors carrying the beta-galactosidase gene (often used for immunological screening). If the cloning event disrupts a normally functional copy of the gene in the vector the resulting plaques would appear clear in the assay. If the phages contain a functional beta-galactosidase gene they will form blue rings around their plaques. Any strain which is not an overproducer of beta- galactosidase will work as indicator host bacteria. - [Read Assay for Phage Containing the Beta-galactosidase Gene Protocol]

BAC-FISH Protocol - http://www.nhm.ac.uk/hosted_sites/schisto/protocols/BAC_FISH.html

Category: BACs Bacterial Artificial Chromosome Protocols

Protocol uses the BIOPRIME reaction kit from GibcoBRL to prepare biotin-labelled BAC DNA which is detected using FITC-Avidin (Vector Labs, DCS grade). Reagents from other manufacturers may work equally well but have not been tested. Includes: Labeling of BAC clones; Ethanol precipitation; Hybridization; Post-hybridisation treatment / detection. - [Read BAC-FISH Protocol]

Buffer Solutions and How they work... - http://www.chemguide.co.uk/physical/acidbaseeqia/buffers.html

Category: Buffers Media, and Solutions: Buffer Theory

This page describes simple acidic and alkaline buffer solutions and explains how they work. - [Read Buffer Solutions and How they work...]

CHROMATIN IMMUNOPRECIPITATION (CHIP) PROTOCOL FOR YEAST PDF - http://research.stowers-institute.org/jeg/2004/cohesin/protocols/ChIP.pdf

Category: DNA Protocols: Chromatin Immunoprecipitation Protocols

Protocol derived from a paper by Miriam Braunstein and is based on work from the Allis lab. - [Read CHROMATIN IMMUNOPRECIPITATION (CHIP) PROTOCOL FOR YEAST PDF]

Dealing with Carryover Contamination in PCR: An Enzymatic Strategy Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4092

Category: PCR Protocols

Protocol for dealing with carryover contamination in PCR- enzymatic strategy. Repeated use of PCR and manipulation of its products cause aerosols that can contaminate neighboring samples and work areas. Such "carryover contamination" can be prevented by including dUTP in place of dTTP for all amplification reactions. - [Read Dealing with Carryover Contamination in PCR: An Enzymatic Strategy Protocol]

First strand cDNA synthesis with Ready-To-Go Beads Protocol - http://www.methods.info/Methods/cDNA/cDNA_ready_to_go.html

Category: PCR Protocols: cDNA Synthesis Protocols

This cDNA synthesis system simplifies your work dramatically. All reaction components are premixed and lyophylised. You have to add your RNA and (for Your-Prime beads) the primer. Another advantage of the system is a little number of pipetting steps required, and therefore reduced risk of Rnase contamination and RNA degradation. - [Read First strand cDNA synthesis with Ready-To-Go Beads Protocol]

General Cell Culture - http://research.cm.utexas.edu/bkitto/Kittolabpage/Protocols/Immunology/GeneralCulture.html

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

Covers general things you need to know to get started in cell culture work. Includes: Sterile Technique; Growth of Your Cell Culture; - [Read General Cell Culture]

Growing Adherent Cells Directly on Tissue Dishes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4290

Category: Cell Culture Protocols

For low-resolution work, cells to be used for staining can be grown directly on regular tissue-culture dishes. It is a convenient method that does not require much preparatory work. - [Read Growing Adherent Cells Directly on Tissue Dishes Protocol]

Home-made Taq Polymerase Purification - http://www.nottingham.ac.uk/~mbzspd/methods/Home_made_Taq_polyme.html

Category: PCR Protocols

PCR polymerase costs can be high. If you are willing to work, you can produce bacteria containing the clone. It appears to produce lots of Taq and is quite stable. The proceedure takes 4 days start to (15 000 units of Taq) finish. The Taq also appears ver - [Read Home-made Taq Polymerase Purification]

How to Work with Slime and Other Cell Wall-Deficient Strains - http://www.fgsc.net/neurosporaprotocols/How%20to%20work%20with%20slime%20final.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to work with slime and other cell wall-deficient strains. - [Read How to Work with Slime and Other Cell Wall-Deficient Strains]

PCR Genotyping from Tail DNA Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/P4.html

Category: Genetics and Genomics: Genotyping Protocols: PCR Genotyping

Protocol for PCR genotyping from tail DNA. This protocol works well for a variety of genes and primer pairs including Tg and KO alleles. Oligonucleotide melting temperatures between 60° and 65° seem to work well. - [Read PCR Genotyping from Tail DNA Protocol]

Preparation of Worm Extracts by Sonication Protocol - http://www.genetics.wustl.edu/tslab/Protocols/Cyto_extract_sonicat_v2.htm

Category: C. elegans Protocols

Protocol was designed to rapidly generate small scale cytosolic extracts of C. elegans for Western or IP (has not been tested for RNA work). The protocol works well for between 50 to 5000 worms and has not been extensively tested on larger a scale, though it should work. Includes: Collection; Sonication; Clearing lysate; Immunoprecipitation. - [Read Preparation of Worm Extracts by Sonication Protocol]

Transient Expression in Protoplasts - http://www.cshprotocols.org/cgi/content/abstract/2007/3/pdb.prot4692

Category: Plant Biology Protocols: Plant Genetics Protocols

The study of transient gene expression provides a useful complement to the study of stably transformed plants. Transient assays offer a quick method of testing the effects of genes, using either phenotypic, molecular, or biochemical readouts. Transient assays based on Agrobacterium-mediated transformation of leaf explants have been described for other plant species, but it is not known how well these assays work in Arabidopsis. - [Read Transient Expression in Protoplasts]

Uterine Transfer Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4380

Category: Mouse Protocols: Mouse Reproductive System Surgery Protocols

Protocol describes a procedure for uterine transfer, which is used for chimera production. The method is based on extensive work which resulted in the first successful development and birth of in-vitro-cultured mouse embryos. It is best to practice this procedure first on a cadaver and then on an anesthetized 2.5-days post coitum (dpc) pseudopregnant mouse using blue Affigel beads rather than embryos. - [Read Uterine Transfer Protocol]

Watching Molecular Motors at Work by Video-Enhanced Light Microscopy - http://www.mih.unibas.ch/Booklet/Booklet96/Chapter2/Chapter2.html

Category: Microscopy Protocols: Optical Microscopy Protocols

The light microscope allows dynamic biological processes to be imaged in their native (i.e., aqueous) environment with relatively high temporal resolution. However, the diffraction-limited resolution is low. When working at or beyond the diffraction-limited resolution of the LM, a disadvantage of fluorescence imaging is the relatively low signal-to-noise (S/N) ratio of the images. However, this can be increased significantly by video and computer technology. - [Read Watching Molecular Motors at Work by Video-Enhanced Light Microscopy]

Yeast RNA Isolation: Small-Scale Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4155

Category: Yeast Protocols: Yeast Nucleic Acid Protocols: Yeast RNA Isolation Protocols

Protocol describes rapid, small-scale yeast RNA isolation. It is based on the work of Schmitt et al. (1990). Note that all containers should be washed in Rnase Away (Invitrogen) or dry baked for 24 hours at 160°C. - [Read Yeast RNA Isolation: Small-Scale 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.

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol. This protocol contains the steps for 3' end rapid amplification of mRNA by PCR. The first-strand cDNA is synthesized from total or poly(A+) RNA by priming from the poly-A tail of the mRNA using a oligo (dT) adaptor primer. The cDNA is then amplified via PCR using a gene-specific primer and an adaptor primer.

Crystal Violet Stain

Crystal Violet stain preparation method and protocol.

Vector Design Protocol for Transgenic Mice Generation

The protocol gives general considerations for the design of targeting vectors for transgenic mice. The protocol shares tips in the design of knock-out and knock-in vectors and some of their strategies for producing homologously recombined embryonic stem cells.

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