2018 Nobel Prize for Medicine or Physiology Awarded to immune checkpoint therapy

Nobel prize for medicine goes to cancer therapy

By Michelle RobertsHealth editor, BBC News online

Two scientists who discovered how to fight cancer using the body's immune system have won the 2018 Nobel Prize for physiology or medicine.

The work, by Professor James P Allison from the US and Professor Tasuku Honjo from Japan, has led to treatments for advanced, deadly skin cancer.

Immune checkpoint therapy has revolutionised cancer treatment, said the prize-giving Swedish Academy.

Experts say it has proved to be "strikingly effective".

Prof Allison, of the University of Texas, and Prof Honjo, of Kyoto University, will share the Nobel prize sum of nine million Swedish kronor - about $1.01 million or 870,000 euros.

Accepting the prize, Tasuku Honjo told reporters: "I want to continue my research ... so that this immune therapy will save more cancer patients than ever."

Prof Allison said: "It's a great, emotional privilege to meet cancer patients who've been successfully treated with immune checkpoint blockade. They are living proof of the power of basic science, of following our urge to learn and to understand how things work."

Treating the untreatable

Our immune system protects us from disease, but it has built-in safeguards to stop it from attacking our own tissue.

Some cancers can take advantage of those "brakes" and dodge the attack too.

Allison and Honjo, now both in their 70s, discovered a way to unleash our immune cells to attack tumours by turning off proteins that put the brakes on.

And that led to the development of new drugs which now offer hope to patients with advanced and previously untreatable cancer.

Immune checkpoint therapy is being used by the NHS to treat people with the most serious form of skin cancer, melanoma.

It doesn't work for everyone, but for some patients it appears to have worked incredibly well, getting rid of the tumour entirely, even after it had started to spread around the body.

Such remarkable results had never been seen before for patients like these.

Doctors have also been using the treatment to help some people with advanced lung cancer.

Prof Charles Swanton, from Cancer Research UK, congratulated the prize winners, saying: "Thanks to this groundbreaking work, our own immune system's innate power against cancer has been realised and harnessed into treatments that continue to save the lives of patients. For cancers such as advanced melanoma, lung, and kidney, these immune-boosting drugs have transformed the outlook for many patients who had run out of options.

"The booming field of immunotherapy that these discoveries have precipitated is still relatively in its infancy, so it's exciting to consider how this research will progress in the future and what new opportunities will arise."

Medicine is the first of the Nobel Prizes awarded each year.

The literature prize will not be handed out this year, after the awarding body was affected by a sexual misconduct scandal.

Recent 10 Noble Prizes for Medicine and Physiology

1. 2018-  James P Allison and Tasuku Honjo for Immune checkpoint therapy to treatments for advanced, deadly cancers.
2. 2017- Jeffrey Hall, Michael Rosbash and Michael Young for unravelling how bodies keep a circadian rhythm or body clock
3. 2016 - Yoshinori Ohsumi for discovering how cells remain healthy by recycling waste.
4. 2015 - William C Campbell, Satoshi Ōmura and Youyou Tu for anti-parasite drug discoveries.
5. 2014 - John O'Keefe, May-Britt Moser and Edvard Moser for discovering the brain's navigating system.
6. 2013 - James Rothman, Randy Schekman, and Thomas Sudhof for their discovery of how cells precisely transport material.
7. 2012 - Two pioneers of stem cell research - John Gurdon and Shinya Yamanaka - were awarded the Nobel after changing adult cells into stem cells.
8. 2011 - Bruce Beutler, Jules Hoffmann and Ralph Steinman shared the prize after revolutionising the understanding of how the body fights infection.
9. 2010 - Robert Edwards for devising the fertility treatment IVF which led to the first "test tube baby" in July 1978.
10. 2009 - Elizabeth Blackburn, Carol Greider and Jack Szostak for finding the telomeres at the ends of chromosomes.

Biodiversity worsens all over the world, in urgency to cope with

An intergovernmental ecological body said on Friday that the biodiversity, the essential variety of life forms on Earth, continued to decline in every region of the world, significantly reducing nature’s capacity to contribute to people’s well-being.

Those alarming trends are endangering economies, livelihoods, food security and the quality of life of people everywhere, according to four peer-reviewed regional reports released by the Intergovernmental Science-Policy Platform on Biodiversity and Ecosystem Services (IPBES).

Human-induced climate change, which affects temperature, precipitation and the nature of extreme events, is increasingly driving biodiversity loss and the reduction of nature’s contributions to people, said Jake Rice, a co-chair of the Americas assessment.

In the Americas, the populations of species are about 31 percent smaller than those was at the time of European colonization, according the report. With the growing effects of climate change added to the other drivers, this loss is projected to reach 40 percent by 2050, it says.

In Africa, by 2100, climate change could also result in the loss of more than half of African bird and mammal species, a 20 to 30 percent decline in the productivity of Africa’s lakes and significant loss of African plant species.

The most recent sad example went to the death of the world’s only remaining male northern white rhino in Kenya on Monday. Its death left only two female northern white rhinos on the planet.

There have been some good news in Asia, however. Over the past 25 years, marine protected areas in the region increased by almost 14 percent and terrestrial protected area by 0.3 percent. Its forest coverage increased by 2.5 percent, with the highest increases in North East Asia (22.9 percent) and by South Asia (5.8 percent).

But the report considered those efforts in Asia insufficient to halt the loss of biodiversity. Unsustainable aquaculture practices, overfishing and destructive harvesting, threaten coastal and marine ecosystems, with projections that, if current fishing practices continue, there will be no exploitable fish stocks in the region by 2048.

Also in Asia, intertidal zones are also rapidly deteriorating due to human activities as up to 90 percent of corals will suffer severe degradation by 2050, even under conservative climate change scenarios.

In the European Union, only 7 percent of marine species and 9 percent of marine habitat types have shown a “favorable conservation status.” Moreover 27 percent of species assessments and 66 percent of habitat types assessments show an “unfavorable conservation status.”

“One of the most important findings across the four IPBES regional assessments is that failure to prioritize policies and actions to stop and reverse biodiversity loss, and the continued degradation of nature’s contributions to people,” said Anne Larigauderie, the Executive Secretary of IPBES.

“Tools like these four regional assessments provide scientific evidence for better decision making and a path we can take forward to achieve the Sustainable Development Goals and harness nature’s power for our collective sustainable future,” said Achim Steiner, Administrator of United Nations Development Program.

Bank of Bioprotocols

Web links of bioprotocols (1)

Web links of bioprotocols (2)

Web Source Protocols for Molecular Biology

1.General Lab Techniques
2. Lab security and basic techniques
3. Advanced lab skills-(1)
4. Advanced lab skills-(2)
5. Molecular Biology
6. DNA isolation & related protocols
7. DNA Purification (glass milk vs electroelution)
8. DNA and RNA sequencing
9. Nucleic acid methods (1)
10.Nucleic acid methods (2)
11.Isolation of DNA,RNA, and Protein simultaneously.
12.DNA mutation detection by SSCP
13.Mouse genotyping by PCR
14.PCR,RT-PCR,Real time PCR etc.
15.Southern blot hybridization
16. Loss of Heterozygosity (LOH)
17.Gene knockout protocol
18.RNA Isolation and Purification
19.Preparation of DNA and RNA probes
20.Northern blot hybridization
21.SiRNA gene knockout
22.Western blot hybridization
23.Molecular cloning
24.Conditional gene transfection(Tet on/off)
25.Protein sequencing
26.Protein labeling techniques
27.Subcellular fractionations
28.EMSA
29.Methylation interference
30.Southwestern blotting
31.Filter binding protocols
32.Footpring protocols
33.DNA (RNA) protein interactions
34.Plasmid and its usefulness
35.DNA library construction
36.Microarray protocols.
37. Protein chips
38.Detecting of protein phosphorylation
39.Protein methods
40.Molecular separation
41.Gene therapy for cancer
42.X-ray crystallography
43.Metabolism and chromatographies
44.GC/MS Background
45.Glucose metabolism and its related protocols
46.PAS staining
47.Polysaccharide sequencing
48.Deoxyribose procedure
49.Ribose metabolism analysis
50.Lactate cleanup and derivative
51.Amino acids
52.Fatty acids
53.Lipid protocols
54.Cholesterol
55.Bile acids
56.Urea procedure
57.Choline Incorporation Assay
58.Isotope Ratio Mass Spectrometer (IRMS)
59.Liquid chromatography / mass spectrometer (LC/MS)
60.Proteomics
61.NMR protocols and tutorials
62.HPLC protocols
63.TLC
64.Molecular labeling
65.Immunology/immunohistochemistry
66.Preparing silanized (+plus) slides
67.Histological fixation
68.Microwave histology
69.Microscopy Techniques
70. Electron microscopy
71.HE staining
72.Nucleic acid stain
73.Special cell & cell fraction stains
74.Antibody purification
75.Antibody storage and handling
76.Conjugation of monoclonal antibodies
77.Antigen retrieval
78.Elisa
79.FRQs for histochemistry
80.Immunoperoxidase staining techniques
81.Immunofluorohistochemistry
82.Confocal microscope technique
83.Laser Capture Microdissection
84.Immunoprecipitation
85.ChIP assay
86.Hybridization in situ
87.Histotechnology--technical methods
88. Flow cytometry (FCM)
89.Kinase assay
90.Cellular Biology
91.General cell culture protocols
92.Chromosome karyotype
93.Proliferation assays (MTT, BrdU, 3H-Thymidine incoporation)
94.Cell cycle assay
95.Migration assay
96.Stem cell & related protocols
97.Apoptosis and related protocols
98.Soft Agar Assay for Colony Formation
99.Aorta ring assay
100.GFP transfection
101.Gene transfection
102.Transformation protocols
103.Blood cell fractionation (white blood cell isolation)
104.Endothelial isolation and culture
105.Isolation and culture human brain microvessel endothelial cells
106.Animal Experiments
107.Blood sampling from animals
108.Basic skills for animal experiments
109.Cancer xenograft animal models
110.Transgenic animal procedures
111.Transgenic cancer models
112.Animal search
113.Animal models for depression-like and anxiety-like behavior
114.Genetics
115.Epigenetics protocols
116.SNPs
117.Genetic analysis
118.Mutagenesis
119.Model Organisms
120.C. Elegans
121.Yeast protocols and resource

Protocols for Molecular Biology

Protocols for DNA
1 Methylene Blue DNA staining protocol
2 Plasmid Protocols--(31-37)
3 Plasmid Protocols--(21-30)
4 Plasmid Protocol--(1-10)
5 Microsatellites Protocols
6 In-Situ hybridization to Embryonic Tissue Section
7 DNA ligation Protocol
8 Colony Hybridization Protocols
9 Colony Hybridization
10 DNA Isolation, Purification and Troubleshooting
11 Deoxyribose Isolation from DNA Degrasion
12 Mitochondrial DNA Isolation
13 Genomic DNA Isolation from Specific Samples
14 Genome DNA Extraction
Protocols for RNA
1 IN VITRO TRANSCRIPTION OF RNA
2 SINGLE CELL mRNA AMPLIFICATION (REVERSE NORTHERN ANALYSIS)
3 Protocols for SiRNA
4 SELEX—in vitro selection
5 Nucleic Acids Research Methods (2)
6 RNA MicroArray Protocol
7 RNA Isolation
Protocols for PCR
1 PCR Based Molecular Cloning
2 Handbook for DNA isolation, RAPD-PCR and PCR-RFLP
3 Introduction to PCR
4 General PCR introduction
5 PCR Animations
6 PCR, RT PCR and Real Time PCR Tutorials
7 PCR Application Manual
8 PCR Elisa
9 PCR RFLP
10 PCR SSCP
11 Real Time PCR
12 RT-PCR Protocols
13 Variants of PCR (2)
14 Variants of PCR (1)
15 AFLP PCR
16 Alu-PCR
17 Asymmetric PCR
18 Colony PCR Protocols
19 Competitive and Quantitative RT-PCR
20 Degenerate PCR
21 Differential Display PCR
22 In Situ PCR
23 Inverse PCR
24 Ligation Mediated Suppression PCR
25 Long PCR Protocols
26 Methylation Specific PCR
27 Multiplex PCR
28 Nested PCR
29 RACE PCR
30 RAPD PCR
31 Rep-PCR
32 TAIL PCR
33 Touchdown PCR
34 Vectorette PCR

Other PCR Web Source (from Google directory)

A Rapid DNA Minipreparation Method Suitable for AFLP and Other PCR Applications - http://pubs.nrc-cnrc.gc.ca/ispmb/ispmb17/17053-1.pdf
Preparation of DNA from plant tissues suitable for PCR methods including AFLP, article by DH CHEN and PC RONALD Department of Plant Pathology, University of California, Davis.

Adjuvants in PCR Reactions - http://info.med.yale.edu/genetics/ward/tavi/p16.html
Brief discussion of additives to improve amplification efficiency and specificity of PCR, by Octavian Henegariu, Yale-New Haven Medical Center.

Amberg Laboratory Protocols: - http://www.upstate.edu/biochem/amberg/protocols.php
Laboratory protocols for PCR work used by the group of David Amberg at the Department of Biochemistry and Molecular Biology Upstate Medical University, Syracuse, New York.

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Anchor Probes for Comparative Mapping of Grass Species - http://greengenes.cit.cornell.edu/anchors/
Article in which probes from different libraries were used to hybridize seven cereals at the Department of Plant Breeding and Biometry, Cornell University, NY.

Attotron Biosensor Corporation - http://www.attotron.com
Research and development company for development of biosensors and related products for the research and educational markets.

BioRad, Amplification, PCR - http://www.bio-rad.com/B2B/BioRad/product/br_category.jsp?
Division of BioRad Laboratories that manufactures and sells instruments for PCR, in Hercules, California, USA.

Degenerate PCR - http://www.dartmouth.edu/~ambros/protocols/other/koelle/degenerate_PCR.html
The identification of novel members of gene families by PCR using degenerate primers is described and protocols given. Article by Michael Koelle 1996 on the web site of Dartmouth College.

Detection of Point Mutations by RFLP of PCR Amplified DNA Sequences - http://www.uni-graz.at/~binder/thesis/node64.html
Thesis abstract about restriction fragment length polymorphism (RFLP) by Alexander Binder 1997.

Detection of Single Nucleotide Mutations in Wheat Using Single Strand Conformation Polymorphism Gels - http://pubs.nrc-cnrc.gc.ca/ispmb/ispmb19/R01-013.pdf
P Martins-Lopez, H Zhang, R Koebner, Plant Mol. Biol. Reporter 19(2001): 159-162. From National Research Coouncil Canada.

DNALC: PCR Animation - http://www.dnalc.org/ddnalc/resources/shockwave/pcranwhole.html
An animation explaining how the Polymerase Chain Reaction (PCR) works, from the Dolan DNA learning center, Cold Spring Harbor Laboratory, USA.

Dolan DNA Learning Centers Gene Almanac - http://www.dnalc.org/home.html
Educational site on topics in genetics and gene expression from Cold Spring Harbor Laboratory, USA.

Effect of PCR Buffer on Multiplex PCR - http://www.qiagen.com/literature/brochures/pcr/pdf/pcrcha03.pdf
Multiplex PCR employs different primer pairs in the same amplification reaction. This requires extensive optimization of annealing conditions. From Quiagen (company).

Fidelity of DNA Polymerases for PCR - http://www.lecb.ncifcrf.gov/~pnh/papers/TIBS/aug95.html
Article by PN Hengen from TIBS 1995

FISH Guide and Troubleshooting - http://info.med.yale.edu/genetics/ward/tavi/FISHguide.html
Links to pages describing influential parameters, with guides on PCR, RT-PCR and multiplex PCR reactions, Taq, FISH, CM-FISH, TM-FISH, microarrays, CCK, slide prep and labeling, maintained by Octavian Henegariu from Yale University, New Haven, CT.

Fluorescence in Situ Hybridisation - http://info.med.yale.edu/genetics/ward/tavi/FISH.html
Technical notes on Fluorescence in situ hybridisation from the Institute of Genetics of Yale School of Medicine, New Haven, Connecticut, USA.

GeneOhm Sciences - http://www.geneohm.com
Tests on group B Streptococcus and methicillin resistant Staphylococcus aureus by PCR / DNA sequencing.

GenHunter - http://www.genhunter.com
Manufacturer of material for differential display PCR in Nashville, Tenn USA.

HiFi DNA - http://www.hifidna.com/
HiFi DNA is a company selling a DNA polymerase for PCR at low temperature giving accurate replication of certain sequences where Taq fails.

Ingenetix GmbH - http://www.ingenetix.com/
Develops technology and products for DNA and mRNA research. Also provide DNA testing for the determination of parentage/paternity and custom DNA sequencing, oligonucleotide synthesis, genotyping services, pharmacogenetics and quantitative PCR, in Vienna, Austria.

Inverse PCR and Cycle Sequencing of P Element Insertions for STS Generation - http://www.fruitfly.org/about/methods/inverse.pcr.html
Step by step protocol, by EJ Rehm, Berkeley Drosophila Genome Project, USA.

Inverse PCR for PAC-end Sequencing - http://www.genetics.wustl.edu/fish_lab/frank/cgi-bin/fish/prot2.html
To generate PCR fragments that contain the ends of PAC inserts that can be sequenced. Protocol by B Barbazuk, Washington University Zebrafish Genome Resources Project, USA.

Inverse PCR for Use with Snyder mTn-lacZ/LEU2-based Mutagenesis - http://labs.fhcrc.org/gottschling/General%20Protocols/ipcr.html
Protocol by M McMurray, Fred Hutchinson Cancer Research Center, Seattle, Wa. USA.

Inverse PCR Protocol - http://www.mcdb.lsa.umich.edu/labs/maddock/protocols/PCR/inverse_pcr_protocol.html
Step by step protocol, from the web site of the Department of Biology, University of Michigan, USA.

Kary B. Mullis - Autobiography - http://nobelprize.org/chemistry/laureates/1993/mullis-autobio.html
The originator of PCR, from the Nobel e-museum web site.

Kary Mullis - http://www.invent.org/hall_of_fame/109.html
Inventor Profile of Kary Mullis, the originator of PCR, from the National Inventors Hall of Fame web site.

Long PCR Protocol - http://twod.med.harvard.edu/labgc/estep/longPCR_protocol.html
Protocol and guidelines for choice of conditions for PCR of long sequences (10 kb or larger). From Genetics Dept., Harvard Medical School, Boston, MA, USA

Nematode ITS1 Size Variation - http://nematode.unl.edu/its_id/EXAMPLES/index.htm
Examples of Restriction Fragment Length Polymorphism (RFLP)electrophoresis slabs for different nematodes, from University of Nebrasca.

Optimizing Multiplex and LA-PCR with Betaine - http://www-lecb.ncifcrf.gov/~pnh/papers/TIBS/jun97.html
LA-PCR = "long and accurate PCR". Article by PN Hengen in TIBS June 1997.

Optimizing PCR Protocols - http://www.jax.org/imr/optimize_pcr.html
Brief guidelines. From the Jackson Laboratory, University of Maine, USA.

PCR Amplification of cDNA Segments by 2 Stage Nested PCR - http://www.ncbi.nlm.nih.gov/SNP/snp_viewTable.cgi?type=method&method_id=555
Protocol from the method database of NIH, USA.

PCR and Multiplex PCR Guide - http://www.info.med.yale.edu/genetics/ward/tavi/Guide.html
Discussions of the parameters influencing the PCR reaction and some PCR and multiplex PCR applications, by Octavian Henegariu on the web site of the Yale - New Haven Medical center.

PCR Animated - http://users.ugent.be/~avierstr/principles/pcrani.html
Animation illustrating the principle of PCR, from the University of Ghent, Belgium.

PCR Gateway - http://www.horizonpress.com/pcr/
A directory of PCR techniques, PCR protocols, PCR troubleshooting, PCR websites and online resources from the publisher Horizon Press.

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PCR Guru - http://www.pcrguru.com/
A downloadable textbook on PCR setup and optimization, not free.

PCR Method Protocols - http://hg.wustl.edu/hdk_lab_manual/pcr/pcrcontents.html
Protocols for PCR posted by the Helen Donis-Keller Laboratory.

PCR Primer Design and Reaction Optimisation - http://web.uct.ac.za/microbiology/pcroptim.htm
Article by Ed Rybicki, Department of Molecular and Cell Biology, University of Cape Town in: Molecular Biology Techniques Manual, on the web site of the University of Cape Town.

PCR Project - http://sunsite.berkeley.edu/biotech/pcr/
Presentations from the University of California at Berkley on PCR, both current research reports and reviews.

PCR Protocol - http://www.mcdb.lsa.umich.edu/labs/maddock/protocols/PCR/general_pcr_protocol.html
Detailed PCR protocol from the web site of the Department of Biology, University of Michigan, USA.

PCR Protocols - http://www.cas.psu.edu/docs/CASDEPT/VET/jackvh/jvhpcr.html
Protocols and technical hints, particularly for reverse transcription PCR, somewhat outdated, compiled by Dr Jack Vanden Heuvel, Department of Veterinary Science and Molecular Toxicology Program, Penn State University

PCR Technology - http://www.accessexcellence.org/LC/SS/PS/PCR/PCR_technology.html
An introduction by Connie Veilleux from the US National Health Museum website.

PCR Troubleshooting - http://info.med.yale.edu/genetics/ward/tavi/Trblesht.html
Limited to conventional straight forward PCR. Page designed and maintained by Octavian Henegariu on the web site of the Yale - New Haven Medical Center.

PCR World - http://pcrworld.blogspot.com
Collection of unreferenced texts on various aspects of PCR.

PCR-ELISA and Related - http://www.btc-bti.com/pcrelisa.htm
P Zhang, CJ Gebhart, D Burden, GE Duhamel: A low technology alternative to real time PCR, technical article on the site of BT&C, Inc Bridgewater, NJ, USA.

Polymerase Chain Reaction - http://www.accessexcellence.com/RC/CT/polymerase_chain_reaction.html
Popular survey article by Mark V. Bloom, DNA Learning Center, Cold Spring Harbor Laboratory, from the web site of the US National Health Museum.

Polymerase Chain Reaction (PCR) - http://www.accessexcellence.org/AB/GG/polymerase.html
A graphic description of the principle of PCR from the US National Health Museum web site.

PrimerDigital - http://primerdigital.com/index.php
International biotechnology company specialized in a design service for PCR primers and probes, PCR-based technology development, projects for development of polymorphism and software development.

Primerfox - http://www.primerfox.com
Free online tool for generation of PCR primers.

Principle of PCR - http://users.ugent.be/~avierstr/principles/pcr.html
Applications in work on aging of Caenorhabditis elegans and phylogeny of nematodes, by Andy Vierstraete, Department of Biology, University of Ghent, Belgium.

Protocols Online: PCR Protocols - http://www.protocol-online.org/prot/Molecular_Biology/PCR/
Extensive collection of PCR protocols and methods from Protocol On Line.

Quantitative PCR Protocol - http://www.jax.org/cyto/quanpcr.html
From the Jackson Laboratory, University of Maine, USA.

RAPD PCR - http://avery.rutgers.edu/WSSP/StudentScholars/project/archives/onions/rapd.html
RAPD stands for Random Amplification of Polymorphic DNA, where the target sequence(s) (to be amplified) is unknown.Brief description, from Rutgers University, USA.

Rational Primer Design Greatly Improves Differential Display-PCR (DD-PCR) - http://nar.oxfordjournals.org/cgi/content/full/25/11/2239
Article: D Graf, AG Fisher, M Merkenschlager: Nucl. Acids Res. 25:11 2239-2240.

Reference in PCR - http://www.gene-quantification.org
Technical aspects of quantitative real-time PCR and RT-PCR. Instruments, kits, dyes, chemistries, and services presented by their manufacturers.

Rep-PCR Genomic Fingerprinting - http://www.msu.edu/user/debruijn/
Bacteria are characterized by Rep-PCR fingerprinting using primers corresponding to naturally occurring repetitive sequences in the interspersed regions.

RFLP Definition - http://vm.cfsan.fda.gov/~frf/rflp.html
RFLP = Restriction Fragment Length Polymorphism, from FDA

Roe Laboratory Protocols - http://www.genome.ou.edu/proto.html
Molecular biological protocols, mostly PCR related used by Bruce A. Roe at the Dept. of Chemistry and Biochemistry, OU, Norman, OK.

Single Tube Confirmation PCR Protocol - http://www-sequence.stanford.edu/group/yeast_deletion_project/single_tube_protocol.html
For characterization colonies of transformed clones of Saccharaomyces, from the web site of the Stanford Genome Technology Center, Palo Alto, CA, USA.

Standard PCR Protocols - http://web.uct.ac.za/microbiology/pcrcond.htm
From Molecular Biology Techniques Manual, from the web site of the University of Cape Town, South Africa.

T-DNA Generated Enhancer Traps in Arabidopsis - http://www.dartmouth.edu/~tjack/
Application of inverse PCR, partial genomic libraries and TAIL-PCR in cloning flanking, at the Department of Biological Sciences, Dartmouth College, Hanover, NH.

Tavi's PCR Protocols - http://info.med.yale.edu/genetics/ward/tavi/PCR.html
A page describing the main parameters and trouble-shooting in PCR. The page is somewhat dated (updated 1997) but still useful.

The PCR Encyclopedia - http://www.pcr-encyclopedia.com/
Describes plans for a free encyclopedia dedicated to the polymerase chain reaction (PCR).

The Web Guide of PCR - http://www.pcrlinks.com/
List of links and forum on the subject and related methodology. Set up and maintained by SJ Krivokapich, National University of Misiones, Argentina.

Thermostable DNA Polymerases - http://arbl.cvmbs.colostate.edu/hbooks/genetics/biotech/enzymes/hotpolys.html
Discussion of their origin and briefly their properties. From the web site of Colorado State University.

Wayward PCR Primers - http://www-lecb.ncifcrf.gov/~pnh/papers/TIBS/jan95.html
Article by PN Hengen from TIBS 1995 on the loss of activity of PCR primers with time.

What the Heck is PCR? - http://people.ku.edu/~jbrown/pcr.html
Popular description of the PCR technique by John C Brown, University of Kansas 1995.

Which DNA Marker for Which Purpose? - http://webdoc.sub.gwdg.de/ebook/y/1999/whichmarker
Compendia of the Research Project "Development, optimisation and validation of molecular tools for assessment of biodiversity in forest trees", European Union DGXII Biotechnology FW IV Research Programme. From the web site of the University Library, Göttingen.

Protocols for Protein
1 Enzyme Assay（43-47）
2 Enzyme Assay（36-42）
3 Enzyme Assay（22-28）
4 Protocols for In Vitro Translation
5 Dot Blot Protocols
6 Selected Protocols of Protein Purification
7 Western Blotting Protocols (1)
Protocols for Cells
1 FACS for Bone marrow macrophages
2 Bone Marrow Macrophages
3 Metabolic Labeling of Cells with 35S
4 Protocols from the science advisory board
5 Introduction to Animal Cell Culture
6 Cell Transfection Protocols
7 Laser Capture Microdissection Protocols
8 Stem Cell Research Techniques and Protocols
9 XTT Cell Viability Assay Protocol
10 Apoptosis Assay Protocols
Protocols for Genetics
1 Genetic Analyses for DNA Protein Interactions
2 Comet Assay
3 Epigenetics Protocols
Protocols for Immunology and IHC
1 Grocott's Methenamine Silver (GMS) stain and troubleshooting
2 Elegans Immunohistochemistry
3 Cytokine Assay Protocols
4 Fluorescence in situ hybridization (FISH)
5 Radioimmunoassay--RIA Protocols
6 Competitive ELISA Protocols
7 Immunohistochemistry Stain for Frozen tissue
Protocols for Chromatography
1 Polysaccharide sequencing
2 Application Guides and Protocols (Nest Group)
3 HPLC Troubleshooting
4 Introduction to HPLC
Protocols for Model Organisms
1 Yeast Protocols
Protocols for Animal Experiment
1 Murine Models of Human Breast Cancer (B)
2 Chorioallantoic Membrane Vascular Assay
3 Guidelines for Techniques with Rodents (Duke University)
4 Tail Vein Injection Techniques
5 Animal Search Engines
Protocols for Basic Techniques
1 H & E Stain Troubleshooting
2 H and E Stain Protocols
3 Electrophoresis Protocols (11-26)
4 Lab. Safety in Life Science