Useful research protocols are right at your fingertips: extensive collections of molecular biology protocols are available online. Even some published print manuals are being posted on the Web. Additionally, many companies supply protocols that are tailored for use with their reagents. The most exciting development in online protocol resources is the increase of laboratory home pages, which provide specific protocols used on the bench.

Most online protocol collections are for molecular biology and recombinant DNA methods. Molecular Biology Protocols, developed by Mark Strom at the Northwest Fisheries Science Center, contains protocols for DNA purification, transformation and sequencing, PCR, electrophoresis, and blotting. A searchable site, it also has a molecular biology techniques forum. Another collection of general molecular biology protocols is the Comprehensive Protocol Collection, maintained by the Ambros Lab at Dartmouth. The site includes protocols for RNA, DNA, and protein methods, and also nematode-related techniques. Both of these collections have been online for about two years, are updated regularly, and feature protocols submitted by other labs.

David Bowtell's extensive collection of biochemistry, molecular biology, and cell biology protocols at the University of Melbourne is an enormous collection that includes everything from mutagenesis and sequencing protocols to cell culture and transfection methods. The methods nicely complement the other molecular biology protocol collections. An online version of the manual Protocols For Recombinant DNA Isolation, Cloning, and Sequencing, edited by Bruce Roe et al. (John Wiley, 1996), links directly to protocols from its table of contents. Promega Corporation Technical Resources provides online access to Promega's technical bulletins and manuals, vector sequences, FAQs, and Promega Notes Magazine. Most protocols are for use with Promega products, but some are general. The Promega site is also searchable.

Several new Web sites provide molecular biology protocol collections. The Whitehead Institute/MIT Center for Genome Research collection contains protocols for DNA purification and sequencing methods. The site also includes protocols for M13 library construction from cosmids, BACs, or P1s. The Frontiers In Bioscience site has tried to assemble a list of the molecular biology protocols from most of the resources listed above, with a few additions.

The majority of the molecular biology protocol collections contain PCR methods. However, several Web sites specialize in PCR procedures. PCR II - A Practical Approach Mutational Analysis: New Mutations by K. Michaelides et al., is excerpted from the book PCR 2: A Practical Approach (Oxford University Press, 1995). The text of the manual has been converted to HTML but contains no internal links. Boehringer Mannheim has made their extensive PCR Applications Manual available for downloading in Adobe Acrobat format. Topics include preparation of PCR templates, PCR protocols, nonradioactive labels in PCR, and in situ PCR. In addition, Gerard R. Lazo's lab site at the Western Regional Research Laboratory, California, has an extensive laboratory protocols list that includes many useful protocols for PCR.

Several Web sites are dedicated to providing protocols for electrophoresis methods. The Molecular Biology Techniques Manual produced at the University of Cape Town contains detailed descriptions and protocols for SDS-PAGE and Western blotting. FMC BioProducts, a supplier of agarose and related reagents, presents technical information that includes protocols for DNA and protein electrophoresis, DNA recovery, and Southern blotting. The ExPASy Molecular Biology Server site at the Geneva University Hospital is an extensive resource for technical information on 2-D PAGE protocols.

The Cell Biology Laboratory Manual, developed by William H. Heidcamp at Gustavus Adolphus College, is a comprehensive resource for cell biology methods. The manual is organized into chapters and includes procedures for cell fractionation, electrophoresis methods, microscopy, and bacterial and fibroblast cell culture. The manual also presents differentiation experiments using C. elegans, D. discoideum, and sea urchins.

For antibody preparation, an excellent online handbook, Antibodies from Design to Assay, is provided at the Perkin-Elmer Web site. Handbook chapters include Designing Antigens, Immunizations, Immunoassays, and Purifying Antibodies. The site provides detailed descriptions, diagrams, and protocols. The Mitchison Lab site also provides protocols and helpful tips for the preparation of antibodies.

Several sources of protocols for cell culture are available online. Bart's Cookbook and Lab Protocols at the Sefton Lab home page contains basic instructions for tissue culture. The Donis-Keller Lab Manual contains many laboratory protocols, but has an especially good section on human cell culture. The Industrial In Vitro Toxicology Group provides a collection of protocols, including liver cell culture and relevant assays.

Several laboratory home pages present immunocytochemical methods. Michael Serfas' lab protocols include basic immunohistochemistry methods and in situ hybridization. Additional immunocytochemistry protocols with good diagrams are provided by the Childs lab at the University of Texas. The Histotechnology Technical Methods site at the University of Nottingham is an extensive general collection of histology protocols.

For Arabidopsis thaliana researchers, two published manuals of protocol collections are available online. Both are available online via the Arabidopsis thaliana Database at Stanford Genomic Resources. Arabidopsis: The Compleat Guide was originally published in 1992 and includes basic plant protocols, tissue culture procedures, DNA and RNA techniques, and microscopy methods. The outstanding Cold Spring Harbor Arabidopsis Molecular Genetics Course Manual from 1996 presents protocols for genetic analysis, in situ hybridization, and gene expression. In addition, Arabidopsis protocols are provided at the home page of the Meyerwitz Lab.

The proliferation of laboratory home pages has allowed researchers to make their lab protocols readily available. (90% of HMS Beagle poll participants use laboratory sites; 63% of HMS Beagle poll participants use their protocols.) These collections are valuable resources for procedures in particular research areas and experimental organisms. In many cases, the online lab protocols contain small details and technical comments that are not available through published research articles.

Many of the yeast labs provide protocol collections. The Herskowitz Lab site includes methods for S. cerevisiae genetics, cell biology, and biochemistry. The Geitz Lab specializes in yeast transformation. The protocols of the Botstein Lab include immunofluorescence and immuno-electron microscopy for yeast. The Forsburg Lab provides protocols for work with the fission yeast S. pombe.

For C. elegans methods, visit the JorgensenLab Protocol Database, which contains general worm procedures, molecular biology, microscopy, and EM protocols. The Carthew Lab Manual is a repository of Drosophila protocols, including somatic mosaic analysis, mutagenesis, and immunostaining. For information on plant methods, the extensive Lazo Laboratory Protocols List provides plant protocols for Arabidopsis, rice, cotton and potato.

Lab Web sites can be good resources for specific experimental techniques. The Mitchison Lab Protocols include protocols for many cytoskeletal techniques in addition to Xenopus oocyte methods. Protocols for the isolation of DNA from mice and genotyping mice by PCR, along with molecular biology and cloning protocols, are provided by the Schimenti Lab. The Faulkner-Jones laboratory presents detailed protocols for in situ hybridization, RNA purification, and eukaryotic DNA analysis. The Spector Laboratory Protocols include immunofluorescence and in situ hybridization of the mammalian cell nucleus.

For recently updated methods information, consult Technical Tips Online, provided by Elsevier Trends Journals. The site presents short, peer-reviewed contributions in a variety of topics and is searchable. Readers can add comments and access other reader's comments about the protocols. Registration is required but is free of charge. The longstanding Bionet newsgroup bionet.molbio.methds-reagnts can be a useful resource for current protocol information. Paul Hengen maintains a list of frequently asked questions from the bionet.molbio.methds-reagnts newsgroup. Newsgroups in specific topic areas can also be sources of technical information. The newsgroups can be accessed via BIOSCI, Deja News or other search engines that support newsgroup searches.

New resources for research protocols are coming online constantly. This article in no way provides a complete list of all the available online protocols. However, hopefully, this article can help you begin to find useful protocols and encourage you to start your own online protocol resource.

REBASE - the Restriction Enzyme Database is an extensive and useful resource for restriction enzyme and methylase information. Restriction enzymes are an integral part of molecular biology and cloning protocols. The REBASE Web site is profiled in the HMS Beagle Site Review.

Protocols on the World Wide Web - a collection of links to different protocol sources, with summaries of each sources' contents.

Cell and Molecular Biology Online - the Methods and Protocols and Current Research pages include links to molecular and cell biology protocol resources and research labs.

LabConsumer - a new section of The Scientist presents cutting-edge information about biological methods and discusses specific research tools used in the laboratory.

The PCR Jump Station - a well-organized collection of links to PCR resources including theory, protocols, and software.

The Antibody Resource Page - includes a compilation of useful antibody Web sites, and resources for locating antibodies.

BBHelp - Molecular Biology Help is a resource for recipes used in molecular biology protocols.

The Laboratory Survival Manual - at the University of Virginia is a resource for information on lab safety and hazardous materials.