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rdf:type |
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lifeskim:mentions |
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pubmed:dateCreated |
2004-7-16
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pubmed:abstractText |
Probe based detection assays form the mainstay of transcript quantification. Problems with these assays include varying hybridization efficiencies of the probes used for transcript quantification and the expense involved. We examined the ability of a standardized competitive RT-PCR (StaRT PCR) assay to quantify transcripts of 4 cell cycle associated genes (RB, E2F1, CDKN2A and PCNA) in two cell lines (T24 & LD419) and compared its efficacy with the established Taqman real time quantitative RT-PCR assay. We also assessed the sensitivity, reproducibility and consistency of StaRT PCR. StaRT PCR assay is based on the incorporation of competitive templates (CT) in precisely standardized quantities along with the native template (NT) in a PCR reaction. This enables transcript quantification by comparing the NT and CT band intensities at the end of the PCR amplification. The CT serves as an ideal internal control. The transcript numbers are expressed as copies per million transcripts of a control gene such as beta-actin (ACTB).
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pubmed:grant |
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pubmed:commentsCorrections |
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Cell Cycle Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Neoplasm,
http://linkedlifedata.com/resource/pubmed/chemical/DNA Probes,
http://linkedlifedata.com/resource/pubmed/chemical/DNA-Binding Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/E2F Transcription Factors,
http://linkedlifedata.com/resource/pubmed/chemical/E2F1 Transcription Factor,
http://linkedlifedata.com/resource/pubmed/chemical/E2F1 protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Proliferating Cell Nuclear Antigen,
http://linkedlifedata.com/resource/pubmed/chemical/RNA, Neoplasm,
http://linkedlifedata.com/resource/pubmed/chemical/Transcription Factors
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pubmed:status |
MEDLINE
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pubmed:month |
Jan
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pubmed:issn |
1476-4598
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pubmed:author |
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pubmed:issnType |
Electronic
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pubmed:day |
23
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pubmed:volume |
3
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
5
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:14741054-Binding, Competitive,
pubmed-meshheading:14741054-Carcinoma, Transitional Cell,
pubmed-meshheading:14741054-Cell Cycle Proteins,
pubmed-meshheading:14741054-Cell Line,
pubmed-meshheading:14741054-Cell Line, Tumor,
pubmed-meshheading:14741054-Computer Systems,
pubmed-meshheading:14741054-DNA, Neoplasm,
pubmed-meshheading:14741054-DNA Probes,
pubmed-meshheading:14741054-DNA-Binding Proteins,
pubmed-meshheading:14741054-E2F Transcription Factors,
pubmed-meshheading:14741054-E2F1 Transcription Factor,
pubmed-meshheading:14741054-Fibroblasts,
pubmed-meshheading:14741054-Gene Expression Regulation, Neoplastic,
pubmed-meshheading:14741054-Genes, Neoplasm,
pubmed-meshheading:14741054-Genes, p16,
pubmed-meshheading:14741054-Humans,
pubmed-meshheading:14741054-Nucleic Acid Amplification Techniques,
pubmed-meshheading:14741054-Proliferating Cell Nuclear Antigen,
pubmed-meshheading:14741054-RNA, Neoplasm,
pubmed-meshheading:14741054-Reproducibility of Results,
pubmed-meshheading:14741054-Reverse Transcriptase Polymerase Chain Reaction,
pubmed-meshheading:14741054-Sensitivity and Specificity,
pubmed-meshheading:14741054-Transcription Factors,
pubmed-meshheading:14741054-Urinary Bladder Neoplasms
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pubmed:year |
2004
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pubmed:articleTitle |
Sensitivity and reproducibility of standardized-competitive RT-PCR for transcript quantification and its comparison with real time RT-PCR.
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pubmed:affiliation |
Department of Pathology, University of Southern California, Keck School of Medicine, 2011 Zonal Ave, HMR 312C, Los Angeles, CA 90033, USA. vpagliarulo@urologia.uniba.it
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pubmed:publicationType |
Journal Article,
Comparative Study,
Research Support, U.S. Gov't, P.H.S.
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