Abstract:
:Multiplexed tandem PCR (MT-PCR) is a process for highly multiplexed gene expression profiling. In the first step, multiple primer pairs are added to the RNA to be analysed together with reverse transcriptase and Taq DNA polymerase. Following reverse transcription, the multiplexed amplicons are simultaneously amplified for a small number of cycles so as to avoid competition between amplicons. The reaction product is then diluted and analysed in multiple individual PCRs using primers nested inside the primers used for the multiplexed amplification. As the second PCR uses a template enriched in the amplicons of interest, the conditions can be optimized to significantly reduce 'primer dimer' formation allowing SYBR Green chemistry to be used for quantification. MT-PCR can be configured for as little as 10 pg RNA (equivalent to a single mammalian cell) and works well with RNA extracted from archival formalin-fixed paraffin-embedded sections. We illustrate MT-PCR with gene expression profiles of breast cancer cell lines.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Stanley KK,Szewczuk Edoi
10.1093/nar/gni182keywords:
subject
Has Abstractpub_date
2005-11-24 00:00:00pages
e180issue
20eissn
0305-1048issn
1362-4962pii
33/20/e180journal_volume
33pub_type
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journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/15.16.6607
更新日期:1987-08-25 00:00:00
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journal_title:Nucleic acids research
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journal_title:Nucleic acids research
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journal_title:Nucleic acids research
pub_type: 杂志文章
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更新日期:2014-08-01 00:00:00
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journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkh179
更新日期:2004-01-12 00:00:00
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journal_title:Nucleic acids research
pub_type: 杂志文章
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journal_title:Nucleic acids research
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journal_title:Nucleic acids research
pub_type: 杂志文章
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更新日期:2014-07-01 00:00:00
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journal_title:Nucleic acids research
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journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/18.24.7235
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journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkm854
更新日期:2008-01-01 00:00:00
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journal_title:Nucleic acids research
pub_type: 杂志文章
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更新日期:1985-09-11 00:00:00
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journal_title:Nucleic acids research
pub_type: 杂志文章
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更新日期:2014-01-01 00:00:00
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journal_title:Nucleic acids research
pub_type: 杂志文章
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journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/8.18.4165
更新日期:1980-09-25 00:00:00
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journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/20.4.645
更新日期:1992-02-25 00:00:00
abstract::Silencing gene expression by siRNAs is rapidly becoming a powerful tool for the genetic analysis of mammalian cells. However, the rapid degradation of siRNA and the limited duration of its action call for an efficient delivery technology. Accordingly, we describe here that Atelocollagen complexed with siRNA is resista...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gnh093
更新日期:2004-07-22 00:00:00
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journal_title:Nucleic acids research
pub_type: 杂志文章
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更新日期:2012-04-01 00:00:00
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journal_title:Nucleic acids research
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journal_title:Nucleic acids research
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journal_title:Nucleic acids research
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journal_title:Nucleic acids research
pub_type: 杂志文章
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journal_title:Nucleic acids research
pub_type: 杂志文章
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更新日期:2008-08-01 00:00:00
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journal_title:Nucleic acids research
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更新日期:2018-02-28 00:00:00
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journal_title:Nucleic acids research
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journal_title:Nucleic acids research
pub_type: 杂志文章
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journal_title:Nucleic acids research
pub_type: 杂志文章
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更新日期:1994-11-11 00:00:00