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Showing posts with label Genomics. Show all posts
Showing posts with label Genomics. Show all posts

Sunday, 24 June 2007

RT PCR Protocols and Analysis

Semiquantitative RT-PCR analysis to assess the expression levels of multiple transcripts from the same sample

Maria Marone1*, Simona Mozzetti1, Daniela De Ritis1, Luca Pierelli1 and Giovanni Scambia1

1 Department of Gynecology and Department of Hematology. Catholic University, L.go A. Gemelli 8, 00168 Rome. Italy.
* To whom correspondence should be addressed: Maria Marone, Department of Gynecology and Department of Hematology. Catholic University, L.go A. Gemelli 8, 00168 Rome. Italy. Email: maria.marone@tiscalinet.it

Link

http://www.biologicalprocedures.com/bpo/arts/1/20/m20.htm

RT-PCR Protocol

Quantitative Reverse Transcription Polymerase Chain Reaction (RT-PCR) and Other PCR Procedures

from

Jack Vanden Heuvel
Penn State University
Department of Veterinary Science
and Molecular Toxicology Program

Link

RT-PCR Protocols ebook

RT-PCR Protocols (Methods in Molecular Biology)

This RT-PCR protocol book describes the detail of novel, useful, and
interesting RT-PCR applications.

protocols available for

highly sensitive detection and quantification of gene expression, the in situ localization of gene,
expression in tissue, and the cloning of genes,

as well as for analyzing T-cell clones and the differential expression of genes,

including laser-capture microdissection (LCM),

real-time and quantitative PCR,

microarray technology, cDNA cloning.

Book Info:
Published in 2002
Published by Humana press
Author Joseph OConnell
ISBN 0896038750  Size 4.30MB

Link

Saturday, 16 June 2007

ENCODE-Encyclopedia Of DNA Elements

ENCODE= the Encyclopedia Of DNA Elements

ENCODE was started in September 2003 to identify all functional elements in the human genome sequence by the National Human Genome Research Institute (NHGRI).

Project has three phases.

1. a pilot project phase-testing and comparing existing methods to rigorously analyze a defined portion of the human genome sequence

2. a technology development
3. a planned production phase

Full details can be found at http://www.genome.gov/10005107

This week Nature published the results of the pilot phase of ENCODE project.


Identification and analysis of functional elements in 1% of the human genome by the ENCODE pilot project
The ENCODE Project Consortium*
http://www.nature.com/nature/journal/v447/n7146/pdf/nature05874.pdf

One of the main conclusions they are pointing to is that there is a lot of places wherethere appears to be conservation of function (transcription and it's regulation) without conservation of sequence. Very interesting.. but long way to go.

Thursday, 14 June 2007

PCR Protocols-Methods in Molecular Biology EBook

PCR Protocols: Current Methods and Applications (Methods in Molecular Biology)

  • Publisher: Humana Press
  • Number Of Pages: 392
  • Publication Date: 1993-01
  • Sales Rank: 914567
  • ISBN / ASIN: 0896032442
  • EAN: 9780896032446
  • Binding: Spiral-bound
  • Manufacturer: Humana Press
  • Studio: Humana Press

    Link

  • Pass: econiches

    Wednesday, 13 June 2007

    Standard PCR Protocol

    Standard PCR Protocols

    This protocol is link to Molecular Biology Techniques Manual
    Third Edition
    Edited by:
    Vernon E Coyne, M Diane James, Sharon J Reid and Edward P Rybicki

    Read Full Protocol

    Overlap Extension PCR

    Overlap Extension PCR is used to create long DNA fragments from short ones.

    or used for Engineering the replication of target DNA through cloning, or changing its genetic code through mutations

    PCR amplify the necessary fragments, using polymerase enzyme. They should have about 15-25 bp overlaps. Use oligo Tm calculators to figure out their annealing temp.
    Clean up or gel extract the correct size band. Use cleaned up fragments as "template". Unlike normal PCR, about 1/2 to 3/4 volume of the extension reaction should be template.
    Use proofreading enzyme for extension. Run 3 reactions of 10,15 and 30 PCR cycles without end primers. (Template extension step) Add end primers, then continue cycling for another 15-20 rounds. Gel extract the correct fragment. Clone into a your desired vector.

    check out the latest Nature Methods Protocol
    http://www.nature.com/nmeth/journal/v4/n5/pdf/nmeth0507-455.pdf

    Monday, 11 June 2007

    Oncogenomics-E Book: Molecular Approaches to Cancer

    Oncogenomics: Molecular Approaches to Cancer

    Cancer is not a single genetic disease but, rather, hundreds of diseases consisting of various combinations of genetic alterations. Many types of genetic alterations contribute to neoplastic transformation. The evidence for this statement has now become common knowledge. In a similar vein, almost everybody believes that genome sequencing is paving the way to a revolution in biology and medicine in general but in oncology in particular. We all mention imatinib and a few monoclonal antibodies when we want to demonstrate that targeted therapies are already more than just a vision. We are, however, still unable to foresee clearly the role

    genomics will play in the prevention, diagnosis, and treatment of cancer in the next 10 or 15 years. This uncertainty stems mainly from a dearth of clinical data, and most of the data we do have are retrospective and therefore most probably biased. As stated in its introduction, this book is probably the first of its kind devoted to the genomics of cancer. I approached it with mixed feelings: on the one hand, I was hoping to find answers to some of my questions, mainly because the span of the book reaches from molecular profiling to model systems for discovering and validating drug targets, and from molecularly targeted pharmacology to clinomics. The latter is a term coined by Daniel Von Hoff et al., whose chapter in this book defines clinomics as the application of genomics to patient care. On the other hand, I approached the book with some
    skepticism, since the field is moving so rapidly that I was afraid to find once again a book that was already too old or just provided reviews that everyone in the field has already read. But I was pleasantly surprised to find a book that is extremely rich in information and detail but is not

    more specialized than necessary. One might criticize certain choices the editors have made -- for instance, providing a discussion of the role of proteomics and genomics in bladder cancer but not in colorectal and lung cancer. This, however, is not very important, since the book was not written for physicians who seek an overview of the latest discoveries in the molecular biology of a specific cancer type. Oncogenomics is very good reading for oncologists who would like to understand what is happening in the laboratory and in the preclinical setting, especially in terms of techniques and research approaches. The book is also worth reading for basic and clinical scientists who intend to plan translational and clinical studies that entail oncogenomics. The essence of the book is to bring together scientists from various specialties and share the same

    language, a prerequisite for the next "leap forward." It seems logical that the book concludes with an essay by A.C. von Eschenbach, the director of the National Cancer Institute, who gives his vision of oncology in 2015. Franco Cavalli, M.D., F.R.C.P.

    Download

    Pass: dmc@lekar

    Friday, 8 June 2007

    Statistical Methods in Bioinformatics: An Introduction-E-Book

    • Hardcover: 588 pages
    • Publisher: Springer; 2 edition (September 30, 2005)
    • Language: English
    • ISBN-10: 0387400826

    Advances in computers and biotechnology have had a profound impact on biomedical research, and as a result complex data sets can now be generated to address extremely complex biological questions. Correspondingly, advances in the statistical methods necessary to analyze such data are following closely behind the advances in data generation methods. The statistical methods required by bioinformatics present many new and difficult problems for the research community.

    This book provides an introduction to some of these new methods. The main biological topics treated include sequence analysis, BLAST, microarray analysis, gene finding, and the analysis of evolutionary processes. The main statistical techniques covered include hypothesis testing and estimation, Poisson processes, Markov models and Hidden Markov models, and multiple testing methods.

    The second edition features new chapters on microarray analysis and on statistical inference, including a discussion of ANOVA, and discussions of the statistical theory of motifs and methods based on the hypergeometric distribution. Much material has been clarified and reorganized.

    The book is written so as to appeal to biologists and computer scientists who wish to know more about the statistical methods of the field, as well as to trained statisticians who wish to become involved with bioinformatics. The earlier chapters introduce the concepts of probability and statistics at an elementary level, but with an emphasis on material relevant to later chapters and often not covered in standard introductory texts. Later chapters should be immediately accessible to the trained statistician. Sufficient mathematical background consists of introductory courses in calculus and linear algebra. The basic biological concepts that are used are explained, or can be understood from the context, and standard mathematical concepts are summarized in an Appendix. Problems are provided at the end of each chapter allowing the reader to develop aspects of the theory outlined in the main text.

    Warren J. Ewens holds the Christopher H. Brown Distinguished Professorship at the University of Pennsylvania. He is the author of two books, Population Genetics and Mathematical Population Genetics. He is a senior editor of Annals of Human Genetics and has served on the editorial boards of Theoretical Population Biology, GENETICS, Proceedings of the Royal Society B and SIAM Journal in Mathematical Biology. He is a fellow of the Royal Society and the Australian Academy of Science.

    Gregory R. Grant is a senior bioinformatics researcher in the University of Pennsylvania Computational Biology and Informatics Laboratory. He obtained his Ph.D. in number theory from the University of Maryland in 1995 and his Masters in Computer Science from the University of Pennsylvania in 1999.

    Comments on the First Edition. "This book would be an ideal text for a postgraduate course.[and] is equally well suited to individual study.. I would recommend the book highly" (Biometrics). "Ewens and Grant have given us a very welcome introduction to what is behind those pretty [graphical user] interfaces" (Naturwissenschaften.). "The authors do an excellent job of presenting the essence of the material without getting bogged down in mathematical details" (Journal. American Staistical. Association). "The authors have restructured classical material to a great extent and the new organization of the different topics is one of the outstanding services of the book" (Metrika).

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    Monday, 4 June 2007

    Cyanine Dye Purification Protocol

     

    Sep-Pak and HPLC purification protocol for Cy dyes (un)incorporated with dNTP Ver. 4.2 (9/18/98)
    compiled by Chih Long Liu, with Kathleen Hayashibara and Linda McAllister; Brown Lab
    [HPLC purification based on dNTP protocol by Suzanne Admiraal, Herschlag Lab]

     

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    Preparation of Fluorescent DNA Probe from HUMAN mRNA or Total RNA using Direct Incorporation (Max Diehn/Ash Alizadeh protocol; 3/15/01)

     

    Preparation of Fluorescent DNA Probe from HUMAN mRNA or Total RNA using Direct Incorporation (Max Diehn/Ash Alizadeh  protocol; 3/15/01)

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    Hybridization of Arrays

    From Brown's Lab

    Last updated: August 10, 1999

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    Post-Processing of Arrays

    From Brown's Lab

    Last updated: September 19, 1999

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    Genomic DNA Labeling Protocol

    From Brown's lab

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    Protocol for Reverse Transcription and Amino-allyl Coupling of RNA

    From Brown's Lab

    The following is a slight modification* of a protocol developed by Joe DeRisi (UCSF) and Rosetta Inpharmatics (Kirkland, WA). Original document can be obtained at www.microarrays.org.

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    Amino-allyl Reverse Transcription Protocol

    From Brown's lab

    (this protocol was adapted by Joe DeRisi from one developed at Rosetta Inpharmatics)

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    how to do Total Bacterial RNA Labeling with Random Hexamers?

    From Brown's Lab

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    Science Protocols