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Monday, May 5, 2008

Chapter 1. General Lab Techniques

Laboratory Manual for Biomedical Research

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Chapter 1. General Lab Techniques


Lab security and basic techniques

Advanced lab skills

Advanced lab skills (2)

Chapter 2. Molecular Biology -- Molecular Separation

Chapter 2. Molecular Biology -- Molecular Separation


Molecular Separation

DNA Agarose Gel Electrophoresis

RNA Agarose Gel Electrophoresis

SDS-Page - Polyacrylamide gel electrophoresis

2D Page-Two - Dimensional Polyacrylamide Gel Electrophoresis

Ion Exchange Chromatography

Gel Filtration Chromatography

Affinity chromatography

Chapter 3. Molecular Biology -- DNA and RNA

Chapter 3. Molecular Biology -- DNA and RNA


Nucleic acid methods (1)

Nucleic acid methods (2)

DNA isolation & related protocols

DNA Purification (glass milk vs electroelution)

DNA, RNA Sequencing

RNA Isolation and Purification

Isolation of DNA,RNA, and Protein simultaneously.

DNA mutation detection by SSCP

Preparation of DNA and RNA probes

Southern blot hybridization

Northern blot hybridization

Loss of Heterozygosity (LOH)

Gene knockout protocol

SiRNA gene knockout

Plasmid and its usefulness

DNA library construction

Microarray protocols.

Basic knowledge of microarray.

Introduction to Microarray.

MicroArray Procedure

Total RNA Isolation from cultured cells.

DNase Treatment of Total RNA

Making the single strand cDNA probe.

Automated Slide Processor (ASP) Version for hybridization.

Washing microarrays in ASP.

Processing of Array slide

Pre-hybridization of the processed slides (NON-Automated version).

Hybridization of Cy3 + Cy5 probe to glass array (NON-Automated version).

Preparation of Dendrimer Cy3 and Cy5.

Washing unbound probe from glass array (NON-Automated version).

Hybridization of Dendrimers (Cy3 and Cy5) to Array (NON-Automated version).

Washing unbound dendrimer from glass array(NON-Automated version).

Microarray Dababases

Troubleshooting

Other Microarray Protocols (1,2)

Gene transfection

Gene therapy for cancer

Molecular cloning

Conditional gene transfection(Tet on/off)

Chapter 4. Genetics

Chapter 4. Genetics


Epigenetics protocols

Mutagenesis protocols

Single nucleotide polymorphisms (SNPs)

Chapter 5. Molecular Biology --PCR Serials

Chapter 5. Molecular Biology --PCR Serials


PCR,RT-PCR,Real time PCR etc.

PCR (General Procedure)

Recommended Reagent Concentrations

Recommended Reaction Conditions

Initial Conditions

Temperature Cycling

"Hot Start" PCR

Asymmetric PCR for ssDNA Production

Detecting Products

Labelling PCR Products with Digoxigenin

Cleaning PCR Products

Sequencing PCR Products

Cloning PCR Products

AND ALWAYS REMEMBER

PCR Primer Design Tools

RT-PCR

Real time PCR

More PCR Protocols Online

Video and Animation of PCR

Mouse Genotyping by PCR

PCR Based Molecular Cloning

PCR Troubleshootings

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.


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.


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.

Chapter 6. Molecular Biology – Protein

Chapter 6. Molecular Biology – Protein


Protein methods

Protein labeling techniques

Protein sequencing

Subcellular fractionations

Western blot hybridization

Western Blot Hybridization Center

Western Blot Full Procedure (from Pierce)

Western Blotting Procedure with Video Show

Protocol 1: Western Blot

Sample protein preparation

Protein concentration assay

Electrophoresis and blotting

Blocking non-specific antigen

Incubation with primary antibody

Incubation with secondary antibody

Substrate (ECL) incubation

Restore a western blot using pierce stripping buffer

Western blot related solutions and buffers

Western blot handbook

Western blot troubleshooting

Protocol 2: Western Blot 2.

Protocol 3: Western Blot 3.

More online Western blot protocols (1) (2)

Protein chips

Kinase assay

Methods for detecting protein phosphorylation

Chapter 7. DNA Protein Interactions

Chapter 7. DNA Protein Interactions


EMSA

ChIP assay

Chapter 8. Immunohistochemistry/immunology

Chapter 8. Immunohistochemistry/immunology


Immunology/Histology

Preparing silanized (+plus) slides

Microscopy Techniques

Confocal microscope technique

Electron microscopy

HE staining

DAPI & PI nucleic acid stain

Hybridization in situ

FISH

Special cell & cell fraction stains

Antibody purification

Antibody storage and handling

Conjugation of monoclonal antibodies

Antigen retrieval

Immunoperoxidase staining techniques

Immunofluorohistochemistry

Immunoprecipitation

Histotechnology--technical methods

FRQs for histochemistry

Laser Capture Microdissection

Elisa

Chapter 9. Cellular Biology

Chapter 9. Cellular Biology


General cell culture protocols

Chromosome karyotype

Proliferation assays (MTT, BrdU, 3H-Thymidine incoporation)

Cell cycle assay

Migration assay

Stem cell & related protocols

Introduction

What are the unique properties of all stem cells?

What are embryonic stem cells?

What are adult stem cells?

What are the similarities and differences between embryonic and adult stem cells?

What are the potential uses of human stem cells and the obstacles that must be overcome before these potential uses will be realized?

Where can I get more information?

International Society for Stem Cell Research (ISSCR)

Culture of Human Embryonic Stem Cells (hESC)

Establishment and characterization of human embryonic stem cell line

Rat stem cell isolation and culture

Mouse stem cell isolation and culture

Induction of Stem Cell Differentiation

Cancer stem cells

Video Data on Stem Cell Research (click to show)

Apoptosis and related protocols

Soft Agar Assay for Colony Formation

Aorta ring assay

GFP transfection

Blood cell fractionation (white blood cell isolation)

Endothelial cell isolation and culture protocols

Flow cytometry (FCM)

Chapter 10. Metabolism, GC/MS, NMR and Proteomics

Chapter 10. Metabolism, GC/MS, NMR and Proteomics


GC/MS Background

Glucose metabolism and its related protocols

PAS staining

Deoxyribose procedure

Ribose metabolism analysis

Polysaccharide sequencing

Lactate cleanup and derivative

Amino acids

Fatty acids

Lipid protocols

Cholesterol

Bile acids

Urea procedure

Choline Incorporation Assay

Isotope Ratio Mass Spectrometer (IRMS)

Liquid chromatography / mass spectrometer (LC/MS)

Proteomics

NMR protocols and tutorials

Chapter 11. Animal Experiments

Chapter 11. Animal Experiments


Blood sampling from animals

Basic skills for animal experiments

Anatomy and physiology of animals

Cancer xenograft animal models

Transgenic animal procedures

Transgenic cancer models

Animal models for depression-like and anxiety-like behavior

Chapter 12. Worm: C. Elegans

Chapter 12. Worm: C. Elegans


C. Elegans Protocols

Chapter 13. HPLC and TLC

Chapter 13. HPLC and TLC


HPLC protocols

TLC

Chapter 14. Buffers and Solutions in Lab.

Chapter 14. Buffers and Solutions in Lab.

Commonly Used Buffer and solution (114 formats in alphabeta)

Compound solutions (Practical Molecular Biology)
How to Make Simple Solutions and Dilutions (Department of Biology, Bates College)
Reto's Buffers&Media Book (Private Page)
Salt solutions(Practical Molecular Biology)
Single Component Solutions (Practical Molecular Biology)
Buffers (Gerard R. Lazo)

Antibiotics (Gerard R. Lazo)
Buffer Listing (Salmon Lab.)

Cell Culture Media and Solutions (Donis-Keller lab)

Commonly used solutions (1)--- 1 ) 2 ) 3 ) 4 ) 5 ) 6 )

Commonly used solutions (2)--- 1 ) 2 ) 3 )

Solutions for molecular cloning---1 ) 2 ) 3 ) 4 ) 5 ) 6 ) 7 ) 8 ) 9 )

Chapter 15. Other Resources

Chapter 15. Other Resources


Free eBooks at Library Online

Cinema Online, Free Movies --(1) (2) (3)

Progresses in Life Science

Progress in Cancer Apoptosis -- (1) (2) (3)

Progress in Gene Therapy --- (1) (2) (3)

Progress in Neuronal Regeneration -- (1) (2) (3)

Progress in AIDS Treatment -- (1) (2) (3)

Progress in Organ Transplantation -- (1) (2) (3)

Progress in Stem Cell Research -- (1) (2) (3)

Free eBooks in life science (1)

Free eBooks in life science (2,3)

Most Updated Biomedical Books

Pathway databases

Pathway Search

Biomedical Job Opportunities

Tools for Statistics

Free Software pDRAW32 to Draw DNA Analysis Charts

Biological Educational Resources

Genetics Education

Resources of Medical Biochemistry

Textbooks and Lab Manuals

Biotech News

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