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Molecular biological protocols, mostly PCR related used by Bruce A. Roe at the Dept. of Chemistry and Biochemistry, OU, Norman, OK.
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Protocols Used in the Roe Laboratory Protocols by keyword or phrase (with Boolean AND/OR/NOT Enter Searchword(s Roe Lab Protocol Book Gel Preparation and ABI 377 XL Protocols updated 3-9-98 Most recent versions of our protocols 384-well DNA Sequencing Template Picking, Growth and Isolation using the QPix...Interface for scanning DNA sequences for restriction sites, without changing encoded protein created. Includes FAQ, table of enzymes and links to tools, maintained at the University of Waterloo, ON, Canada.
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Redirecting to watcut/uwaterloo.ca The site you are looking for has been replaced by a new version that can be found at http watcut.uwaterloo.ca...Links to pages describing influential parameters, with guides on PCR, RT-PCR and multiplex PCR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.
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Tavi's FISH protocols and slide preparation guide Page designed and maintained by Octavian Henegariu (Email: or WARNING: The information provided in these pages is copyrighted and is intended for individual use only. No parts of this work (text, tables or pictures) may be commercialized, published or otherwise...Detailed protocol for terminal restriction fragment length polymorphism (T-RFLP), which allows the fingerprinting of a community by analyzing the polymorphism of a certain gene. Article by V Grüntzig, B Stres, HL Ayala del RÃo and JM Tiedje, Center for Microbial Ecology, Michigan State University, USA
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RDP 404 Alert The page you were seeking is no longer available Return to the RDP-II Release 9 Site OR Return to the RDP-II Release 8.1 Site 160; If you have immediate questions, please contact rdpstaff@msu.edu directly or you may use our hotline...List of links and forum on the subject and related methodology. Set up and maintained by SJ Krivokapich, National University of Misiones, Argentina.
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PCR Links.com -The Web Guide Of Polymerase Chain Reaction Technique PCR R firm aims to cut lab time for diagnostics Posted By: Labtechnologist.com Published: Friday, 26-11-2004 ·U.S. Patent Issued To Applera For Real-Time PCR Posted By: LIMSfinder.com Published: Wenesday, 10-11-2004 ·Research for faster Polymerase Chain Reaction technology Posted By:News-Medical.Net Published: Monday,...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
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Long PCR Protocol R results from the use of two polymerases: a non-proofreading polymerase is the main polymerase in the reaction, and a proofreading polymerase (3 to 5 exo) is present at a lower concentration. Following the results of Cheng et al 1)...An animation explaining how the Polymerase Chain Reaction (PCR) works, from the Dolan DNA learning center, Cold Spring Harbor Laboratory.
http://www.dnalc.org/shockwave/pcranwhole.html
HiFi DNA is a company selling a DNA polymerase for PCR at low temperature giving accurate replication of certain sequences where Taq fails.
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HiFi DNA: High Fidelity DNA Polymerase for Low Temperature PCR R technology to the next level with the following unique features: Cycling temperatures between 37C and 70C, using a processive DNA polymerase with a truly proof-reading function. Extends primers over the GC-rich regions where the heat-resistant DNA polymerases like Pfu*...Presentations from the University of California at Berkley on PCR, both current research reports and reviews.
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PCR Project What is the polymerase chain reaction (PCR It is the supreme biotechnological invention. PCR has transformed molecular biology through vastly extending the capacity to identify, manipulate and reproduce DNA. It makes abundant what was once scarce the genetic material required...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
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PCR Protocols Please visit their web sight to order: http www.crcpress.com/catalog/3344.htm An important new manual for those interested in PCR techniques, new applications, and troubleshooting existing reactions Description Molecular toxicology is an emerging discipline that utilizes molecular and cell biology to understand...An introduction by Connie Veilleux from the US National Health Museum website.
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PCR Technology Polymerase chain reaction (PCR) has rapidly become one of the most widely used techniques in molecular biology and for good reason: it is a rapid, inexpensive and simple means of producing relatively large numbers of copies of DNA molecules from...A popular description of the way it works, its importance in different connections, legal aspects. Authored by TM Powledge at the Federation of the American Societies of Experimental Biology.
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The Polymerase Chain Reaction A series of articles for general audiences The Polymerase Chain Reaction By Tabitha M. Powledge It is hard to exaggerate the impact of the polymerase chain reaction. PCR, the quick, easy method for generating unlimited copies of any fragment of...A graphic description of the principle of PCR from the US National Health Museum web site.
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302 Found The document has moved here. Apache/2.0.40 Server at www.accessexcellence.org Port 80...Applications in work on aging of Caenorhabditis elegans and phylogeny of nematodes, by Andy Vierstraete, Department of Biology, University of Ghent, Belgium.
http://allserv.rug.ac.be/~avierstr/principles/pcr.html
Bacteria are characterized by Rep-PCR fingerprinting using primers corresponding to naturally occurring repetitive sequences in the interspersed regions.
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Rep-PCR Genomic Fingerprinting Home Page The utility of a recently developed method to classify bacteria on the basis of their genomic fingerprint patterns was investigated, using collections of both symbiotic and pathogenic plant-associated bacteria. The genomic fingerprinting method employed is based on the use of...A page describing the main parameters and trouble-shooting in PCR. The page is somewhat dated (updated 1997) but still useful.
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Tavi's PCR protocols Page designed and maintained by Octavian Henegariu (Email: or WARNING: The information provided in these pages is copyrighted and is intended for individual use only. No parts of this work (text, tables or pictures) may be commercialized, published or otherwise...Intended for specialists planning PCR procedures in their laboratories. From the University of Cape Town updated 2001.
http://web.uct.ac.za/microbiology/pcr.htm
Popular description of the PCR technique by John C Brown, University of Kansas 1995.
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What the Heck is PCR? Polymerase chain reaction (PCR) is a technique which is used to amplify the number of copies of a specific region of DNA, in order to produce enough DNA to be adequately tested. This technique can be used to identify with...Popular survey article by Mark V. Bloom, DNA Learning Center, Cold Spring Harbor Laboratory, from the web site of the US National Health Museum.
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Polymerase Chain Reaction Pasteur once remarked that chance favors the prepared mind and certainly the history of scientific progress supports his contention. The annals of science provide numerous examples of serendipitous discovery. Some are mythic, such as Newton's discovery of gravity following his...Inventor Profile of Kary Mullis, the originator of PCR, from the National Inventors Hall of Fame web site.
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Invent Now Hall of Fame Search Inventor Profile The polymerase chain reaction, which was devised by Kary Mullis, has revolutionized DNA technology. PCR amplifies specific DNA sequences from very small amounts of complex genetic material. The amplification produces an almost unlimited number of highly purified DNA molecules suitable...Article by PN Hengen from TIBS 1995 on the loss of activity of PCR primers with time.
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Methods and reagents is a unique monthly column that highlights current discussions in the newsgroup bionet.molbio.methds-reagnts, available on the Internet. This month's column discusses the failure of PCR primers that have been kept a long time to amplify DNA. For...Article by PN Hengen from TIBS 1995
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Methods and reagents is a unique monthly column that highlights current discussions in the newsgroup bionet.molbio.methds-reagnts, available on the Internet. This month's column discusses the fidelity of polymerases used for amplification of DNA by the polymerase chain reaction. For details...Brief protocols for PCR in use at the Genome Sequencing Center of the Washington University in St Louis Medical School.
http://genome.wustl.edu/tools/protocols/index.php?fin4=1
Article by TD Brock from the web site of Department of Bacteriology, University of Wisconsin - Madison on the thermophilic organisms in Yellowstone hot springs, including Thermus aquaticus from which Taq polymerase was obtained.
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Life at High TemperaturesLife at High Temperatures The thermophilic bacteria that live in the Yellowstone hot springs have been the foundation of impressive developments in medicine and biotechnology. The unique thermostable enzymes of these bacteria are finding wide industrial and medical use, and have become the basis...Discussion of their origin and briefly their properties. From the web site of Colorado State University.
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Thermostable DNA Polymerases It is interesting how some seemingly esoteric or obscure discoveries can, years later, be catapulted to something of immense practical importance. Such is the history of Taq DNA polymerase. The original report of this enzyme, purified from the hot springs...LA-PCR = "long and accurate PCR". Article by PN Hengen in TIBS June 1997.
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Methods and reagents is a unique monthly column that highlights current discussions in the newsgroup bionet.molbio.methds-reagnts, available on the Internet. This month's column discusses the use of additives for optimizing the amount and quality of product obtained through multiplex and...Brief discussion of additives to improve amplification efficiency and specificity of PCR, by Octavian Henegariu, Yale-New Haven Medical Center.
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Adjuvants Page designed and maintained by Octavian Henegariu (Email: or WARNING: The information provided in these pages is copyrighted and is intended for individual use only. No parts of this work (text, tables or pictures) may be commercialized, published or otherwise...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.
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Guide.html Page designed and maintained by Octavian Henegariu (Email: or As I am currently pursuing a new junior faculty position, the Yale URL and email may change in the near future. If messages sent to the yale.edu address bounce back to...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.
http://web.uct.ac.za/microbiology/pcroptim.htm
Limited to conventional straight forward PCR. Page designed and maintained by Octavian Henegariu on the web site of the Yale - New Haven Medical Center.
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PCR Troubleshooting Page designed and maintained by Octavian Henegariu (Email: or As I am currently pursuing a new junior faculty position, the Yale URL and email may change in the near future. If messages sent to the yale.edu address bounce back to...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.
http://webdoc.sub.gwdg.de/ebook/y/1999/whichmarker
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.
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%PDF-1.2 17 0 obj endobj xref 17 34 0000000016 00000 n 0000001027 00000 n 0000001290 00000 n 0000001497 00000 n 0000001652 00000 n 0000001673 00000 n 0000002474 00000 n 0000002580 00000 n 0000002687 00000 n 0000004036 00000 n 0000004144 00000...In asymmetric PCR, the low concentration primer is quantitatively incorporated into double stranded DNA after an appropriate number of cycles. If this primer is fluorescent labelled, the dsDNA can be quantitated from the fluorescence.
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FA-PCR and FCS by Kinjo R products in homogeneous solution by fluorescence correlation spectroscopy Masataka Kinjo Laboratory of Biophysics, Research Institute for Electronic Science, Hokkaido University, 060, Sapporo, JAPAN Tel 81-11-706-2890 Fax 81-11-706-4964 e-mail: kinjo@imd.es.hokudai.ac.jp Keywords: asymmetric PCR; FCS; fluorescent probe; fluorescence measurement, PCR; quantitative...A report and protocol, from Fondation Jean Dausset.
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Alu-PCR hybridization This library was divided in 64 superpools of 4 microtiter plates each. Then each superpool (384 clones) was represented by 22 pools: 8 pools of half-plates (the first pool contains all half of the first plate, the second pool contains...From Molecular Biology Techniques Manual, from the web site of the University of Cape Town, South Africa.
http://web.uct.ac.za/microbiology/pcrcond.htm
For characterization colonies of transformed clones of Saccharaomyces, from the web site of the Stanford Genome Technology Center, Palo Alto, CA, USA.
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Untitled Document R protocol has been designed to be performed in individual reaction tubes. We usually test three colonies from each transformation along with a wild-type control. A series of five different PCR tests are performed on each colony to confirm both...Article in which probes from different libraries were used to hybridize seven cereals at the Department of Plant Breeding and Biometry, Cornell University, NY.
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Anchor Probes for Comparative Mapping of Grass Species Comparative mapping using DNA proves can be used to combine genetic information from these related species. One of the prerequisites for comparative mapping is a genetic linkage map for each species. RFLP maps presently exist for rice (Oryza sativa L...Detailed PCR protocol from the web site of the Department of Biology, University of Michigan, USA.
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General PCR Protocol R in different size reaction volumes and in smaller tubes as long as they fit in the thermocycler. Materials: template DNA (genomic, plasmid, cosmid, bacterial/yeast colony, etc primers (resuspended to a known concentration with sterile TE) buffer (usually 10X, usually...step by step protocol, from the web site of the Department of Biology, University of Michigan, USA.
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Inverse PCR protocol Isolate partially purified chromosomal DNA using a modification of the Magic MiniPreps DNA Purification System from Promega (see atta ched) as follows Spin down 500 µl of an overnight culture in a 1.5 ml microfuge tube Resuspend in 200 µl...Protocol for RAP-PCR to study prokaryotic gene expression, from the Enterococcus Research Site, University of Oklahoma, USA.
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The Enterococcus Research Site Random Arbitrarily Primed PCR A generalized protocol for RAP-PCR to study prokaryotic gene expression Please see the following reference for additional details and discussion: Shepard, B. D and M. S. Gilmore. 1999. Identification of aerobically and anaerobically induced genes in Enterococcus faecalis by random...Article: D Graf, AG Fisher, M Merkenschlager: Nucl. Acids Res. 25:11 2239-2240.
http://nar.oupjournals.org/cgi/content/full/25/11/2239
Manufacturer of material for differential display PCR in Nashville, Tenn USA.
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Differential Display: GenHunter Online Japan identified a new adipocytokine, visfatin, that is highly enriched in the abdominal fat versus subcutaneous fat of both humans and mice (Science. 2005, 307:426-430 They showed that Visfatin level in plasma increases during the development of obesity. Most importantly,...An introduction with principle, applications and description of the technique. From the web site of Kebangsaan University, Malaysia.
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Degenerate PCR Primers which have a number of options at several positions in the sequence so as to allow annealing to and amplification of a variety of related sequences. eg: 5’-TCG AAT TCI CCY AAY TGR CCN T-3’ Y pYrimidines C T...Step by step protocol, by EJ Rehm, Berkeley Drosophila Genome Project, USA.
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For recovery of sequences flanking PZ, PlacW and PEP elementsFor SuPorP, pGT1, and p{Epgy2} elements, see the Bellen lab web site [HERE] E. Jay Rehm Berkeley Drosophila Genome Project I. Quick Fly Genomic DNA Prep Standard fly mini prep (30...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.
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WashU-Zfish Project Radiation Hybrid (RH) Maps R fragments that contain the ends of PAC inserts that can be sequenced. For inverse PCR we cut the PAC once in the vector (near outward primer site) and once at unknown site in insert (and many other places outside...Protocol by M McMurray, Fred Hutchinson Cancer Research Center, Seattle, Wa. USA.
http://www.fhcrc.org/labs/gottschling/misc/ipcr.html
Application of inverse PCR, partial genomic libraries and TAIL-PCR in cloning flanking, at the Department of Biological Sciences, Dartmouth College, Hanover, NH.
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T-DNA enhancer traps in Arabidopsis How the lines are generated Collecting seeds for distribution to the ABRC stock center Detailed information about the lines pD991 enhancer trap vector Sample Patterns Cloning flanking DNA Overview of the project This project involved the generation of more than...Company promoting diagnostic tests for genetic analysis; rolling circle amplification technology (RCAT), developed for amplifying signals from DNA, RNA and proteins, from Westport, CT.
http://www.molecularstaging.com/Pages/RCAToverview_.html
Multiplex PCR employs different primer pairs in the same amplification reaction. This requires extensive optimization of annealing conditions. From Quiagen (company)
http://www.qiagen.com/literature/brochures/pcr/pdf/pcrcha03.pdf
This method works because the poison allows the formation of deletion products but titers out full-sized products. From the Biotechnology Laboratory, University of British Columbia.
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Poison Primer 1 R protocols to detect rare deletions in DNA from complex populations of mutagenized worms typically depend on a large size differential between the wild-type product and the deletion product. These protocols are used with libraries in which deletion representation is...Protocol from the method database of NIH, USA.
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Method Detail Submitted method description: PCR amplification of cDNA segments by 2 stage nested PCR protocol Protocol: NESTED PCR Template: 50 ng cDNA Primer: each 0.5 uM dNTPs: each 0.2 mM PCR Buffer: 5 ul [10X, 100 mM Tris-HCl (pH 8.4 500...P Zhang, CJ Gebhart, D Burden, GE Duhamel: Molecular and Cellular Probes, 14(2) 101-108, on the site of BT&C, Inc.
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PCR-ELISA, an alternative to real time PCR for detecting products of the polymerase chain reaction A less expensive alternative to real time PCR BTC/OPS Diagnostics 672 Rt. 202-206 North Bridgewater, NJ TEL 908) 253-3444 FAX 908) 575-1660 Email:info@btc-bti.com HOME About BTC Opportunities Management Contact Us PRODUCTS Grinding Products Shakers Vortexers Freeze Drying Custom Products NEW...Protocols including standard nested PCR, from the Biotechnology Laboratory, University of British Columbia.
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Vancouver Lab Home Laboratory is part of an international consortium, working to produce null mutations in all identified and predicted genes in the C. elegans genome. Targeted gene inactivation is accomplished using a PCR-based reverse-genetics method. Knockout mutations have been generated by the...RFLP = Restriction Fragment Length Polymorphism, from FDA
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RFLP Definition If two organisms differ in the distance between sites of cleavage of a particular restriction endonuclease, the length of the fragments produced will differ when the DNA is digested with a restriction enzyme. The similarity of the patterns generated can...Examples of Restriction Fragment Length Polymorphism (RFLP)electrophoresis slabs for different nematodes.
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Nematode ITS1 Size Variation Examples of PCR-RFLP for Nematode Diagnostics Nematode ITS1 Size Variation ITS1 Size Polymorphism 11 Tylenchid Genera Nematode rDNA ITS1 Size Variation PCR-RFLP Variation Among Several Cyst Nematode Species PCR-RFLP Variation Among Several Cyst Nematode Species PP Presentation PCR-RFLP Variation Between...RFLP = Restriction Fragment Length Polymorphism. By Alexander Binder 1997.
http://www-ang.kfunigraz.ac.at/~binder/thesis/node64.html
Illustration of principle. CAPS corresponds to RFLP. From Corvallis Forestry Research Community, Corvallis, USA.
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JFIF C 0-(0 C b 1AV 2QSU BTaf 8q3Ru 4FHb 57CDG 6E‚r 1AQRa 2q SáBb#r 3C ho_ u Cz k E x 81 s O LPÒ»T^z qS t 9 UÌE»U*jz Qj OyµÜnV R gE kv 9`ÙtGDPj ZYRf p K sy...Discussion of the technique and variations, literature references. By S McQuaid, Institute of Child Health, University College London, UK.
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CMGS-SSCP/21.10.97 Slowly but surely towards better scanning for Mutations Richard Cotton, TIG February 1997, Vol. 13, No. 2, pp 43-46 Mutation Scanning Considerations Features of the ideal mutation scanning method able to scan kilobase lengths 100% detection rate no false positives...P Martins-Lopez, H Zhang, R Koebner, Plant Mol. Biol. Reporter 19(2001): 159-162. From National Research Coouncil Canada.
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%PDF-1.3 20 0 obj endobj xref 20 20 0000000016 00000 n 0000000764 00000 n 0000000819 00000 n 0000001205 00000 n 0000001385 00000 n 0000001519 00000 n 0000001703 00000 n 0000001810 00000 n 0000001918 00000 n 0000003219 00000 n 0000003325 00000...This technique is used to obtain the 3'end of a cDNA. Protocol by R Hastings, Biochemistry, University of Nottingham Medical School, UK.
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3 RACE PCR This technique is used to obtain the 3'end of a cDNA, it requires some sequence information internal to the mRNA under study. The sequence information obtained from this technique can be utilised to obtain full length cDNA clones using the...RAPD stands for Random Amplification of Polymorphic DNA, where the target sequence(s) (to be amplified) is unknown.Brief description, from Rutgers University, USA.
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RAPD PCR A which are essentially unknown to the scientist (random Often, PCR is used to amplify a known sequence of DNA. Thus, the scientists chooses the sequence he or she wants to amplify, then designs and makes primers which will anneal...Brief description of Random Amplified Polymorphic DNA, from Oklahoma State University, USA.
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RAPD's Molecular polymorphisms can be identified at random and used for genetic mapping. Facts The complexity of eukaryotic nuclear DNA is sufficiently high that by chance pairs of sites complementary to single octa- or decanucleotides may exist in the correct orientation...Protocol for Randomly Amplified Polymorphic DNA, by H Judelson, University of California, Riverside, USA.
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RAPD analysis J research machine using microliter plates; double times for Perkin Elmer 480 machine using standard 0.5 ml tubes) 94 C, 15 sec. 1X 94 C, 30 sec; 35 C, 30 sec; 72 C, 60 sec. 36 X 72 C, 2.5...Animation illustrating the principle of PCR, from the University of Ghent, Belgium.
http://allserv.rug.ac.be/~avierstr/principles/pcrani.html
Manufactures and sells products for molecular biology including thermal cyclers and sequencers / genotypers.
http://www.mjr.com/
R & D company for development of biosensors and related products for the research and educational markets.
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Attotron Biosensor Corporation Attotron is a new company devoted to research and development of new technologies for low-cost biosensors and related products for the research and educational markets. What does the company name mean? The name comes from the metric prefix Atto meaning...Extensive collection of PCR protocols and methods from Protocol On Line.
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Molecular Biology/PCR Protocols The reaction is cycled involving template denaturation, primer annealing, and the extension of the annealed primers by DNA polymerase until enough copies are made for further analysis. Subcategories Bisulfite Conversion Based PCR (6) Degenerate PCR (4) In Situ PCR (6)...From the Jackson Laboratory, University of Maine, USA.
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Quantitative PCR Protocol The Jackson Laboratory Dn mice is based on doing simultaneous quantitative PCR amplification of a gene or genes in the Ts65Dn chromosome and a control gene on another chromosome (in this case Apob) and comparing the average change (delta) in threshold cycle (CT)...Brief guidelines. From the Jackson Laboratory, University of Maine, USA.
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Optimizing PCR Protocols The Jackson Laboratory This can be the buffer provided with the DNA polymerase.)1.5 mM MgCl2 final (optimal may need to be determined, range: 1.0-5.0 mM)0.2 mM dNTPs final0.4 M each primer final (range: 0.2-1.0 M)0.25-0.5 units thermostable DNA polymerase per reaction Melting temperature...Notes about PCR applications, principles, research results, from Roche.
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Innovative Tools for Amplification ...56 page booklet on oligonucleotides, PCR and RT-PCR, from Biosource (company).
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%PDF-1.3 155 0 obj endobj xref 155 36 0000000016 00000 n 0000001071 00000 n 0000001788 00000 n 0000002008 00000 n 0000002220 00000 n 0000002770 00000 n 0000003502 00000 n 0000003543 00000 n 0000003566 00000 n 0000004735 00000 n 0000004757 00000...Rapid Amplification of cDNA Ends. Protocol, from National Institute of Genetics, Japan.
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EARLY EMBRYOGENESIS/MIDLINE SIGNALING Preparation and handling of poly A+ RNA 1. General precautions The integrity and purity of your poly A+ RNA used as starting material is an important element of high-quality cDNA synthesis. The following precautions will help you avoid contamination and...Protocols for PCR posted by the Helen Donis-Keller Laboratory.
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PCR Methods Polymerase Chain Reaction Purification of Synthetic Pligos by Ion Exchange Chromatography Purification and Sequencing of PCR Product Back to Main Table of Contents...The originator of PCR, from the Nobel e-museum web site.
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Kary B. Mullis Autobiography Carolina in the foothills of the Blue Ridge Mountains. My dad's family had a general store, which I never saw. My grandparents on his side had already died before I started noticing things. My mother's parents were close to me...Technical aspects of quantitative real-time PCR and RT-PCR. Instruments, kits, dyes, chemistries, and services presented by their manufacturers.
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Gene-Quantification.com Gene-Quantification.info The reference in qPCR Academic Industrial Information Platform N web page illustrates the usefulness of reliable quantification strategies, e.g. absolute relative-quantification assays, and the difference between them in kinetic PCR kinetic RT-PCR. RT-PCR is the technique of choice for analyzing mRNA in extremely low abundance. But real-time RT-PCR...Basic discussion of the optimization of PCR, from the web site of Brinkmann Instruments.
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Eppendorf North America Optimizing DNA Amplification Protocols using the Eppendorf Mastercycler This method was first described in 1985 (1) and enables selective in vitro amplification of a special DNA fragment, thereby emulating the cellular in vivo DNA replication. In spite of the method's basically simple operation, often enough it is not...Article on the influence of the design of the primer on the outcome of PCR, by Dr Vincent R. Prezioso, published by Eppendorf, North America.
http://www.eppendorfna.com/appli...tions/pcr_appl_primer.asp?SX=NF&
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.
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degenerate_PCR_2.html The identification of novel members of gene families by PCR using degenerate primers has been considered more of an art than a science, so much so that the methods books I've come across have been too timid to discuss the...Educational site on topics in genetics and gene expression from Cold Spring Harbor Laboratory, USA.
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Dolan DNA Learning Center's Gene Almanac COLD SPRING HARBOR LABORATORY. ALL RIGHTS RESERVED. EMAIL DOLAN DNA LEARNING CENTER Problems? Go to our text-only page....Tests on gruup B Streptococcus and methicillin resistant Staphylococcus aureus by PCR / DNA sequencing.
http://www.geneohm.com
Applications in work on aging of Caenorhabditis elegans and phylogeny of nematodes, by Andy Vierstraete, Department of Biology, University of Ghent, Belgium.
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Principle of the PCR This is necessary to have enough starting template for sequencing. The cycling reactions There are three major steps in a PCR, which are repeated for 30 or 40 cycles. This is done on an automated cycler, which can heat and...Animation illustrating the principle of PCR, from the University of Ghent, Belgium.
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PCR animated (last update 20-08-1999) Figure 6 Animated picture of PCR. back to principle of the PCR back to homepage...