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Protocols
- 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: 4923, Reviews: 0 Write review Cached - Quantitative RT-PCR
(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: 12218, Reviews: 0 Write review - 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: 8878, 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: 7431, 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: 2459, 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: 3167, 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: 3360, Reviews: 0 Write review Cached