Abstract:
:Quantification of nucleic acids, especially of mRNA, is increasingly important in biomedical research. The recently developed quantitative real-time polymerase chain reaction (PCR) - a highly sensitive technology for the rapid, accurate and reproducible quantification of gene expression - offers major advantages over conventional quantitative PCR. Transcript quantification is performed in the exponential phase of the PCR reaction through extrapolation of fluorescence signals from a standard calibration curve which represents the initial copy number for a given fluorescence signal. We have developed a method for gene transcript quantification which is based on a LightCycler - assisted real-time PCR in combination with a simple and rapid approach for the construction of external cRNA standards with identical gene sequences as the target gene. Synthesis of cRNAs was performed by in vitro transcription with T7 RNA polymerase followed by reverse transcription and real-time PCR. We applied this approach for transcript quantification of eukaryotic initiation factor 3 p110 (EIF3S8) mRNA in normal testicular tissue. We also present a rapid and simple strategy for the construction of cRNA standards for use in real-time PCR.
journal_name
Mol Cell Probesjournal_title
Molecular and cellular probesauthors
Fronhoffs S,Totzke G,Stier S,Wernert N,Rothe M,Brüning T,Koch B,Sachinidis A,Vetter H,Ko Ydoi
10.1006/mcpr.2002.0405subject
Has Abstractpub_date
2002-04-01 00:00:00pages
99-110issue
2eissn
0890-8508issn
1096-1194pii
S0890850802904056journal_volume
16pub_type
杂志文章abstract::An assay for rapid detection of herpes simplex virus in infected cells is described. The assay utilizes in situ hybridization with photobiotin-labelled double-stranded DNA probes prepared from HSV-1 DNA cloned in plasmid vectors. The assay provided an alternative method for earlier detection of virus in cell cultures ...
journal_title:Molecular and cellular probes
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abstract::A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay targeting the nucleocapsid phosphoprotein gene of infectious bronchitis virus (IBV) was developed. The detection limits for the IBV RT-LAMP assay were 10(1) 50% egg infection dose (EID(50)) per 50 microl of titrated viruses and no cross-rea...
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journal_title:Molecular and cellular probes
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journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1006/mcpr.1994.1025
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journal_title:Molecular and cellular probes
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journal_title:Molecular and cellular probes
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doi:10.1016/j.mcp.2008.10.001
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journal_title:Molecular and cellular probes
pub_type: 杂志文章
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journal_title:Molecular and cellular probes
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journal_title:Molecular and cellular probes
pub_type: 杂志文章
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journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1006/mcpr.1994.1069
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journal_title:Molecular and cellular probes
pub_type: 杂志文章
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journal_title:Molecular and cellular probes
pub_type: 杂志文章
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journal_title:Molecular and cellular probes
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journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2011.06.001
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journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2006.12.001
更新日期:2007-06-01 00:00:00
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journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2010.02.001
更新日期:2010-08-01 00:00:00
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journal_title:Molecular and cellular probes
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doi:10.1016/0890-8508(89)90026-1
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journal_title:Molecular and cellular probes
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journal_title:Molecular and cellular probes
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doi:10.1016/j.mcp.2008.09.004
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journal_title:Molecular and cellular probes
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journal_title:Molecular and cellular probes
pub_type: 杂志文章
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journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2014.09.003
更新日期:2015-02-01 00:00:00
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journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2008.02.001
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journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2004.11.008
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journal_title:Molecular and cellular probes
pub_type: 杂志文章
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journal_title:Molecular and cellular probes
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doi:10.1016/j.mcp.2005.12.006
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journal_title:Molecular and cellular probes
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journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/0890-8508(92)90055-3
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abstract::Non-small-cell lung carcinoma (NSCLC) accounts for approximately 80% of lung cancers with a high metastatic potential. Elucidating the mechanism of NSCLC metastasis will provide new promising targets for NSCLC therapy and benefit its prognosis. Plasmacytoma variant translocation 1 (PVT1) has been proven to be overexpr...
journal_title:Molecular and cellular probes
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