Detection of PRL-2 gene expression in hepatocellular carcinoma by real-time fluorescence quantitative PCR

2009 ◽  
Vol 8 (4) ◽  
pp. 210-213
Author(s):  
Chao Cheng ◽  
Amos Ela Bella ◽  
Ailin Guo ◽  
Guoyong Wu ◽  
Weikang Wu
Keyword(s):  
Gene Expression ◽  
Hepatocellular Carcinoma ◽  
Real Time ◽  
Quantitative Pcr ◽  
Fluorescence Quantitative Pcr

A Real-Time Continuous Flow Polymerase Chain Reactor for DNA Expression Quantification

2007 ◽  
Author(s):  
Michael B. Sayers ◽  
Tara M. Dalton
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Quantitative Pcr ◽  
Real Time Pcr ◽  
Continuous Flow ◽  
Quantitative Polymerase Chain Reaction ◽  
Ccd Camera ◽  
Fluorescent Imaging ◽  
Intensity Level ◽  
Polymerase Chain

Real-time quantitative Polymerase Chain Reaction (PCR) is an extremely sensitive and reliable method for quantifying gene expression, allowing subtle shifts in gene expression to be easily monitored. Currently, stationary real-time PCR is readily achieved using fluorescent labels which increase in fluorescence as the DNA is exponentially amplified. Quantitative PCR is used in a myriad of applications. However currently most commercial real-time PCR devices are batch process stationary well based systems, limiting their throughput. Continuous flow microfluidic PCR devices have allowed for advancement in terms of improved PCR throughput and reduced reagent usage. As part of an overall total analysis system a device integrating all the functional steps of continuous flow realtime quantitative PCR has been designed and fabricated. Initially the PCR reaction mixture is segmented into nano-litre PCR reactors which are then thermally cycled on a two temperature fifty cycle flow-through PCR device, which allows laser induced fluorescent imaging of the nanoreactors. Previous studies into continuous flow PCR have demonstrated endpoint fluorescent measurements, however this research allows PCR nanoreactors to be fluorescently monitored after every PCR thermal cycle. Fluorescent optical monitoring is achieved through laser excitation of the nanoreactors while a Charged Coupled Device (CCD) camera is used to record the fluorescent emissions from the nanoreactors. Intensity analysis of the recorded images is then preformed using MATLAB to accurately determine the fluorescence intensity level, thereby allowing real-time quantitative amplification curves to be generated. This has major advantages over existing continuous flow PCR devices which use endpoint fluorescence and capillary electrophoresis, as the amplification curves allow far more information to be gleaned and allow the initial DNA template concentration to be accurately determined.

Human disease characterization: real-time quantitative PCR analysis of gene expression

2001 ◽  
Vol 6 (20) ◽  
pp. 1062-1067 ◽  
Author(s):  
James V Snider ◽  
Mark A Wechser ◽  
Izidore S Lossos
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Quantitative Pcr ◽  
Human Disease ◽  
Pcr Analysis ◽  
Real Time Quantitative Pcr

Validation of reference genes for accurate normalization of gene expression for real time-quantitative PCR in strawberry fruits using different cultivars and osmotic stresses

Gene ◽  
2015 ◽  
Vol 554 (2) ◽  
pp. 205-214 ◽  
Author(s):  
Vanessa Galli ◽  
Joyce Moura Borowski ◽  
Ellen Cristina Perin ◽  
Rafael da Silva Messias ◽  
Julia Labonde ◽  
...  
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Quantitative Pcr ◽  
Reference Genes ◽  
Real Time Quantitative Pcr ◽  
Accurate Normalization

scholarly journals Establishing reference genes for use in real-time quantitative PCR analysis of early equine embryos

2011 ◽  
Vol 23 (2) ◽  
pp. 353 ◽  
Author(s):  
Damien B. B. P. Paris ◽  
Ewart W. Kuijk ◽  
Bernard A. J. Roelen ◽  
Tom A. E. Stout
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Quantitative Pcr ◽  
Embryo Development ◽  
Reference Genes ◽  
Stable Expression ◽  
Pcr Analysis ◽  
Blastocyst Development ◽  
Real Time Quantitative Pcr

Real-time quantitative PCR (qPCR) is invaluable for investigating changes in gene expression during early development, since it can be performed on the limited quantities of mRNA contained in individual embryos. However, the reliability of this method depends on the use of validated stably expressed reference genes for accurate data normalisation. The aim of the present study was to identify and validate a set of reference genes suitable for studying gene expression during equine embryo development. The stable expression of four carefully selected reference genes and one developmentally regulated gene was examined by qPCR in equine in vivo embryos from morula to expanded blastocyst stage. SRP14, RPL4 and PGK1 were identified by geNorm analysis as stably expressed reference genes suitable for data normalisation. RPL13A expression was less stable and changed significantly during the period of development examined, rendering it unsuitable as a reference gene. As anticipated, CDX2 expression increased significantly during embryo development, supporting its possible role in trophectoderm specification in the horse. In summary, it was demonstrated that evidence-based selection of potential reference genes can reduce the number needed to validate stable expression in an experimental system; this is particularly useful when dealing with tissues that yield small amounts of mRNA. SRP14, RPL4 and PGK1 are stable reference genes suitable for normalising expression for genes of interest during in vivo morula to expanded blastocyst development of horse embryos.

Sign in / Sign up

Export Citation Format

Share Document