URT-PCR Designed for Human Tissue and mRNA Amplification and Quantitation
- 技术优势
- URT for all tested transcriptsmRNA specific PCRStandard RNA isolation techniques are acceptable – no additional purification stepsMultiple transcripts can be amplified from one total RNA sampleStandard housekeeper reference transcripts (i.e. β-actin, GAPDH) are valid controlsUsable with standard RT-PCR strategies/kits – only primers differQuantitation of low copy transcripts in small tissue samples is feasibleSensitivity similar to standard RT-PCR protocols
- 技术应用
- Gene expression assays, from traditional to microarraysQuantitative RT-PCR applicationsGene regulation researchGene expression phenotyping of cells and tissues for research or clinical molecular pathologySmall sample (e.g. micro dissection) or single cell expression assaysInter-individual expression phenotyping for disease susceptibility
- 详细技术说明
- The system has been readily employed in the common and currently used qualitative and real-time quantitative PCR protocols assessing gene expression in small samples of human tissues. The strategy has therefore proven to be universally applicable to all tested transcripts in all tested gene-expression assay contexts.
- *Abstract
-
Researchers have devised a novel reverse transcription strategy that utilizes a mRNA-specific RT primer designed to be universal for all mRNA transcripts. The employment of the mRNA-specific RT strategy obviates the limitations posed by total RNA preps contaminated with gDNA. Subsequent PCR is performed integrating features of the universal RT primer design, and is not complicated by the presence of gDNA nor pseudogene. Messager RNA can thus be amplified very specifically in the setting of gDNA contamination, without further steps. Purification of total RNA to mRNA becomes unnecessary, a particularly critical advantage when handling small tissue samples, as in human tissues. Both DNase treatment after RNA isolation, and no-RT controls in the PCR step to assess the presence of pseudogene, become unnecessary. Standard reference “housekeeper” transcripts such as β-actin and GAPDH, otherwise contaminated by gDNA-encoding pseudogenes, can be amplified specifically and quantitatively. Multiple transcripts can be amplified from the same total RNA aliquot, maximizing the efficiency of reverse transcription on limited RNA samples.
- *IP Issue Date
- Nov 28, 2006
- *IP Type
- Utility
- *Principal Investigation
-
Name: Simon D. Spivack, Ph.D. Gregory J. Hurteau
Department:
- 国家
- United States
- 附加资料
- Inventor: ZHENG, Bojian | SUI, Hongyan | LIN, Yongping
Priority Number: WO2010066112A1
IPC Current: C12N0015113 | A61K004800 | A61P003114
Assignee Applicant: The University of Hong Kong
Title: siRNA COMPOSITIONS AND METHODS FOR POTENTLY INHIBITING VIRAL INFECTION | COMPOSITIONS D'ARNSI ET PROCÉDÉS POUR INHIBER FORTEMENT UNE INFECTION VIRALE | COMPOSITIONS D’ARNSI ET PROCÉDÉS POUR INHIBER FORTEMENT UNE INFECTION VIRALE
Usefulness: siRNA COMPOSITIONS AND METHODS FOR POTENTLY INHIBITING VIRAL INFECTION | COMPOSITIONS D'ARNSI ET PROCÉDÉS POUR INHIBER FORTEMENT UNE INFECTION VIRALE | COMPOSITIONS D’ARNSI ET PROCÉDÉS POUR INHIBER FORTEMENT UNE INFECTION VIRALE
Summary: For preparing an agent for stimulating interleukin-6 (IL-6) production or for stimulating β -defensin production in a subject; for screening of potential antiviral agents; and as antiviral agents (all claimed), for prophylaxis or treatment of viral infection, preferably by H5N1 influenza virus.
Novelty: New small interfering RNAs useful for prophylaxis or treatment of influenza A virus infection, comprise pentanucleotide motif or reverse motif having specific nucleotide sequence
- 主要类别
- 生物医学
- 细分类别
- 病原
- 申请号码
- 7,141,372
- 国家/地区
- 美国
