Here's how to do it

The spider’s web

Here’s how to do it

If you haven’t got the right protocol,it could be right at your fingertips.Extensive collections of molecularbiology protocols are availableonline. In addition, many companiessupply protocols that are tailored foruse with their reagents. But the mostexciting development in onlineprotocol resources is the proliferationof laboratory home pages thatprovide specific lab protocols.

Molecular Biology Protocols, atthe Northwest Fisheries ScienceCenter, and the ComprehensiveProtocol Collection, maintained bythe Ambros Lab at Dartmouth,continue to be excellent resourcesfor molecular biology methods. Thesites include protocols for RNA,DNA and protein methods, areupdated regularly and featureprotocols submitted by other labs.

David Bowtell’s SignalTransduction Group at the Universityof Melbourne provides an enormouscollection of protocols that includes

everything from mutagenesis andsequencing to cell culture andtransfection. The methods nicelycomplement the other molecularbiology protocol collections. TheWhitehead Institute–MIT Center forGenome Research collection containsprotocols for DNA purification andsequencing methods and for M13library construction from cosmids,BACs or P1s. In addition, the manualProtocols for Recombinant DNAIsolation, Cloning and Sequencing isavailable online.

There are several web sites thatspecialize in PCR procedures. PCR II— A Practical Approach MutationalAnalysis: New Mutations isexcerpted from the book PCR 2: APractical Approach published byOxford University Press. BoehringerMannheim has made the extensivePCR Applications Manual available fordownloading in Adobe Acrobatformat. Topics include preparation ofPCR templates, PCR protocols, non-radioactive labels in PCR and in situPCR. The lab site of Gerard Lazohas an extensive Laboratory ProtocolsList that includes many for PCR.

Several web sites are dedicated toprotocols for electrophoresis

methods. The Molecular BiologyTechniques Manual containsdetailed descriptions and protocolsfor SDS-PAGE, western blotting andPCR. FMC BioProducts, a supplierof agarose and related reagents,presents technical information thatincludes protocols for DNA andprotein electrophoresis, DNArecovery and Southern blotting. TheExPASy Molecular Biology Serversite is an extensive resource for two-dimensional PAGE protocols andtechnical information.

For antibody preparation, thePerkin–Elmer site provides anexcellent online handbook withdetailed descriptions, diagrams andprotocols: Antibodies from Design toAssay. The Mitchison Lab site alsoprovides protocols and helpful tipsfor the preparation of antibodies.

There are several online sourcesof protocols for cell culture. Bart’sCookbook and Lab Protocols at theSefton Lab home page contains basicinstructions for tissue culture. TheDonis–Keller Lab Manual containsmany laboratory protocols, but has anespecially nice section on human cellculture. The Industrial In VitroToxicology Group provides acollection of protocols, includingliver cell culture and relevant assays.

Several laboratory home pagespresent immunocytochemicalmethods. Michael Serfas’ LabProtocols include basicimmunohistochemistry methods andin situ hybridization. Additionalimmunocytochemistry protocols withnice diagrams are provided by theChilds lab. The HistotechnologyTechnical Methods site at theUniversity of Nottingham is anextensive general collection ofhistology protocols.

New resources for researchprotocols are coming onlineconstantly. In particular, specific labprotocols can provide the sort ofhandy hints that make the differencebetween the success and failure of anexperiment.

Address: spider@cursci.co.uk

Magazine R739

Molecular Biology Protocols http://research.nwfsc.noaa.gov/protocols.htmlComprehensive Protocol Collection http://www.dartmouth.edu/artsci/bio/ambros/protocols.htmlHome page for the Melbourne Signal Transduction Group

http://grimwade.biochem.unimelb.edu.au/sigtrans.htmlWhitehead Institute–MIT Center for Genome Research

http://www.genome.wi.mit.edu/sequencing/protocols/protocols.htmlProtocols For Recombinant DNA Isolation, Cloning and Sequencing

http://www.genome.ou.edu/protocol_book/protocol_index.htmlPCR II - A Practical Approach Mutational Analysis: New Mutations

http://146.179.66.63/usr/WWW/WebPages/database.dir/methods.dir/pcrpract.htmBoehringer Mannheim PCR Applications Manual

http://biochem.boehringer.com/prod_inf/manuals/pcr_man/pcr_toc.htmlLaboratory Protocols List http://wheat.pw.usda.gov/homepage/lazo/methods/Molecular Biology Techniques Manual http://www.uct.ac.za/microbiology/manualin.htmFMC BioProducts Technical Information http://www.bioproducts.com/technical/Technical information on 2-D PAGE http://expasy.hcuge.ch/ch2d/technical-info.htmlAntibodies: From Design to Assay http://www2.perkin-elmer.com/pa/340913/340913.htmlMitchison Lab Protocols http://skye.med.harvard.edu/Protocols/protocols.htmlBart’s Cookbook and Lab Protocols

http://carmen.salk.edu/users/Sefton_web/Hyper_protocols/TableOfContents.htmlDonis–Keller Lab Manual http://hdklab.wustl.edu/lab_manual/index.htmlIndustrial In Vitro Toxicology Group http://www.invitro.org/tests.htmMichael Serfas’ Lab Protocols http://icarus.uic.edu/~azathoth/protocols.htmlOur Favorite Cytochemical Protocols http://cellbio.utmb.edu/childs/cytochem.htmHistotechnology Technical Methods http://www.ccc.nottingham.ac.uk/~mpzjlowe/pathprot.html

This month’s URLs