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  • Standard RT-PCR (Pamela Stanley Lab Wiki, Yale University)
    RT-PCR or reverse transcription PCR refers to PCR that uses product of an RT reaction as template. In effect, the PCR amplifies cDNA fragments. In one-tube RT-PCR, RNA and PCR primers are added to a reaction mix that is thermocycled for RT first followed by for PCR. One-tube RT-PCR reaction mixes are supplied by many manufacturers. Drawbacks to their use include lack of flexibility. Otherwise, RT-PCR is a two step process.
    http://stanxterm.aecom.yu.edu/wiki/index.php?page=...
    Added: Fri Mar 20 2009, Hits: 17638, Reviews: 0  Write review   Cached
  • RT-PCR procedures (Roche Diagnostics Corporation)
    RT-PCR can be performed as either a two-step (one tube or two tube) or an one-step (coupled RT-PCR) procedure. This protocol introduces both methods and their advantages.
    http://www.roche-applied-science.com/pcr/applicati...
    Added: Sat Sep 21 2002, Hits: 1445, Reviews: 0  Write review   Cached
  • overviews on quantitative kinetic RT-PCR PDF file (Dr. Michael W. Pfaffl, Gene Quantification )
    This review discusses the technical aspects involved, contrasts conventional and kinetic RT-PCR methods for quantitating gene expression and compares the different kinetic RT-PCR systems. It illustrates the usefulness of these assays by demonstrating the significantly different levels of transcription between individuals of the housekeeping gene family, glyceraldehyde-3-phosphate-dehydrogenase (GAPDH).
    http://www.wzw.tum.de/gene-quantification/adope-pd...
    Added: Sun Sep 22 2002, Hits: 7506, Reviews: 1 Read reviews  Write review  
  • Preparation of Template RNA for RT-PCR (Roche Diagnostics Corporation)
    Guidelines for preparing high quality RNA template for RT-PCR.
    http://www.roche-applied-science.com/pcr/applicati...
    Added: Sat Sep 21 2002, Hits: 779, Reviews: 0  Write review   Cached
  • Competitive Quantitative RT-PCR (Ambion)
    Detailed protocol for competitive quantitative RT-PCR
    http://www.ambion.com/techlib/prot/bp_1760.pdf
    Added: Tue May 14 2002, Hits: 2337, Reviews: 0  Write review  
  • RNA Extraction Protocol of LCM tissue and RT-PCR for mRNA analysis (NIH Laser Capture Microdissection Core Lab)
    http://dir.nichd.nih.gov/lcm/LCMTAP.htm#Sample RNA...
    Added: Tue May 14 2002, Hits: 2456, Reviews: 0  Write review  
  • Competitive RT-PCR (Dieter Kaufmann)
    For quantifying mRNA, internal standard RNAs are added in a defined quantity to the RNA sample prior to the RT reaction. The resulting standard cDNA is coamplified with the same primers as the endogenous target sequence. Its PCR product is approximately 50 nucleotides smaller. This method allows measurement of small differences (as low as factor 2) in mRNA amount between RNA samples.
    http://www.uni-ulm.de/~dkaufman/rt-pcr.html
    Added: Tue May 14 2002, Hits: 4689, Reviews: 0  Write review   Cached
  • RT Reaction and RT-PCR (Cowin Lab, New York University Medical Center)
    http://celljunctions.med.nyu.edu/lab/notprivate/me...
    Added: Mon Mar 16 2009, Hits: 4944, Reviews: 0  Write review   Cached
  • Semiquantitative RT-PCR analysis to assess the expression levels of multiple transcripts from the sa (Biological Procedures Online )
    a semiquantitative RT-PCR protocol optimized to extract RNA from as little as 10,000 cells and to measure the expression levels of several target mRNAs from each sample
    http://www.biologicalprocedures.com/bpo/arts/1/20/...
    Added: Wed Sep 25 2002, Hits: 3212, Reviews: 0  Write review   Cached
  • Protocol for Real-Time RT-PCR (Dr. Xiaowei Wang at the Department of Molecular Biology, Massachusetts General Hospital)
    This protocol describes the detailed experimental procedure for real-time RT-PCR using SYBR Green I. The procedure begins with reverse transcription of total RNA. The cDNA is then used as template for real-time PCR with gene specific primers. You may need to modify this protocol if you use different reagents or instruments for real-time PCR.
    http://pga.mgh.harvard.edu/primerbank/PCR_protocol...
    Added: Fri May 14 2004, Hits: 2091, Reviews: 0  Write review   Cached
  • Detecting DNA Contamination in RT-PCR (Qiagen)
    To avoid Detecting DNA contamination in RT-PCR, use controls, design approporiate primers.
    http://www1.qiagen.com/resources/info/detecting_dn...
    Added: Sun Jul 21 2002, Hits: 3579, Reviews: 0  Write review   Cached
  • RT-PCR Primer Design (Roche Diagnostics Corporation)
    How to design RT-PCR primers allowing differentiation between the amplified product of cDNA and an amplified product derived from contaminating genomic DNA.
    http://www.roche-applied-science.com/pcr/applicati...
    Added: Sat Sep 21 2002, Hits: 893, Reviews: 0  Write review   Cached
  • Nested RT-PCR From Paraffin Section (Hans Popper)
    This protocol is for the non-isotopic detection of hepatitis C RNA and albumin mRNA (as an internal control) from 4 micron sections of formalin-fixed, paraffin-embedded liver biopsies by RT-PCR. The procedure for RNA extraction from formalin fixed paraffin embedded section and PCR amplification is described.
    http://zapruder.pds.med.umich.edu/users/Frank/HepC...
    Added: Tue May 14 2002, Hits: 299, Reviews: 0  Write review   Cached
  • Quantitative RT-PCR PDF file (The Morimoto Lab, Northwestern University)
    This 9-page protocol describes every aspects of quantitative RT-PCR in great detail, including the principles, RNA preparation, primer design, preparation of internal control...
    http://www.biochem.northwestern.edu/morimoto/resea...
    Added: Sun Mar 28 2004, Hits: 11741, Reviews: 0  Write review  
  • Semi -Quantitative RT-PCR (Pamela Stanley Lab Wiki, Yale University)
    The RT-PCR method can be used not only to detect specific mRNAs but also to semi-quantitate their levels. Thus, one can compare levels of transcripts in different samples. This can be done in two different ways. One is to quantitate against levels of transcripts from a control, house-keeping gene (such as actin and GAPDH). (Transcription of house-keeping genes is believed to be unaffected by almost all experimental conditions.) The second method is to add an exogenous, primer-specific PCR template during PCR.
    http://stanxterm.aecom.yu.edu/wiki/index.php?page=...
    Added: Fri Mar 20 2009, Hits: 7085, Reviews: 0  Write review   Cached
  • Quantitative RT-PCR and Other PCR Procedures (Jack Vanden Heuvel, Penn State University)
    A must-read protocol for PCR beginners as well as verterans. It touches in depth aspects specific to quantitative RT-PCR as well as those common to standard PCR.
    http://www.cas.psu.edu/docs/CASDEPT/VET/jackvh/jvh...
    Added: Sun Mar 28 2004, Hits: 8290, Reviews: 0  Write review  
  • Semi-Quantitative RT-PCR (Mike A. Dyer)
    From RNA isolation, reverse transcription to PCR...
    http://axon.med.harvard.edu/~cepko/protocol/mike/R...
    Added: Tue May 14 2002, Hits: 3158, Reviews: 0  Write review   Cached
  • Gene-specific RT-PCR PDF file (Molecular Profiling Initiative, NCI)
    This protocol is used for microdissected RNA, but may apply to other situations. It introduce three three priming methods for generating first strand cDNA from mRNA: 1) Oligo (dT)-based priming is preferred when the RNA is of high quality, since first strand polymerization should be continuous from the 3' end of the transcript to the desired sequence regardless of the length of the 3' UTR. 2)Random hexamer priming is recommended for fragmented mRNA (lengths <500 bases)where there may be discontinuity between the desired sequence and the poly A tail. 3)A primer that is specific to the gene of interest is best when amplifying low abundance transcripts.
    http://cgap-mf.nih.gov/Protocols/PDFFormat/GeneSpe...
    Added: Sat Aug 24 2002, Hits: 12576, Reviews: 0  Write review  
  • How can I treat my RNA sample before RT-PCR to eliminate DNA contamination? (Invitrogen)
    http://invitrogen.custhelp.com/cgi-bin/invitrogen....
    Added: Tue May 14 2002, Hits: 1792, Reviews: 0  Write review   Cached
  • Reverse Ligation Mediated RT-PCR (Antisense Research Group)
    RL-PCR can be broken down into a number of simple steps. Synthesis by in vitro transcription and purification of an RNA linker species. Extraction of total RNA from cells into which oligodeoxynucleotide has been delivered by, for example, streptolysin O permeabilization. Ligation of the RNA linker to all available 5' monophosphates in the purified total cellular RNA sample. Reverse Transcription of the RNA using a gene - specific primer. Amplification of specific fragments by PCR using linker - specific and gene - specific primers. Sub- amplification of the first PCR product using linker specific and nested, labelled, gene specific primers.
    http://www.liv.ac.uk/~giles/methods/rlpcr.htm
    Added: Tue May 14 2002, Hits: 1532, Reviews: 0  Write review  
  • Gene-specific RT-PCR (Cancer Gene Anatomy Project, NCI)
    Protocol may be used for amplifying individual transcripts from RNA recovered from microdissected cell populations. The procedure for reverse transcription is as regular RT, but for PCR, radioactive dCTP is used and PCR products are separated by running polyacrylamide gel.
    http://cgap-mf.nih.gov/Protocols/DNARNAProteomicAn...
    Added: Tue May 14 2002, Hits: 666, Reviews: 0  Write review   Cached
  • FAQs on Real-Time RT-PCR (Dr. Xiaowei Wang at the Department of Molecular Biology, Massachusetts General Hospital)
    Some questions and answers on real-time PCR primer design and amplification
    http://pga.mgh.harvard.edu/primerbank/faq.html
    Added: Fri May 14 2004, Hits: 1067, Reviews: 0  Write review   Cached
  • DNA Contamination in RT-PCR (Ambion)
    Present data showing levels of DNA contamination in RNA generated by different procedures and suggest several precautionary measures which can be implemented to reduce the impact of this persistent problem.
    http://www.ambion.com/techlib/tb/tb_176.html
    Added: Tue May 14 2002, Hits: 2814, Reviews: 0  Write review   Cached
  • RT-PCR (Lazo Lab)
    First strand cDNA synthesis and PCR amplification
    http://wheat.pw.usda.gov/~lazo/methods/lazo/revtrp...
    Added: Tue May 14 2002, Hits: 3887, Reviews: 0  Write review   Cached
  • How can I tell if my RT-PCR product in RNA specific? (Invitrogen)
    http://invitrogen.custhelp.com/cgi-bin/invitrogen....
    Added: Tue May 14 2002, Hits: 2082, Reviews: 0  Write review   Cached

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