qPCR NEWS - June 2011 - focus on RNA integrity
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Dear researcher,
dear Gene Quantification page reader,
Our newsletter informs about the latest news in quantitative real-time PCR (qPCR and qRT-PCR), which are compiled and summarised on the Gene Quantification homepage. The focus of this newsletter issue is:
- Update - RNA integrity - http://RNA-integrity.gene-quantification.info
- qPCR 2011 eConference - http://eConference.qPCR2011.net
- US qPCR Symposium in Santa Clara, CA - 1st-4th November 2011 - http://EVENTS.gene-quantification.info
- MIQE qPCR APP for iPhone, iPad and iPod - iOS Universal - http://MIQE.gene-quantification.info
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http://RNA-integrity.gene-quantification.info
It is generally known that total RNA quality has a distinct influence on the validity and reliability of quantitative PCR results. In addition, the recently published MIQE guidelines focus on the pre-PCR steps and state the importance of RNA quality assessment. Various studies showed the impairing effect of ongoing RNA degradation on mRNA expression results. Therefore, the verification of RNA integrity prior to downstream applications like RT-qPCR and mircroarrays is indispensable. A fast and reliable assessment of RNA integrity can be done with the Eukaryote Total RNA Nano Assay of the Agilent 2100 Bioanalyzer. The importance of RNA quality should also be considered in new applications such as the investigation of miRNA expression profiles. With the Agilent Small RNA Assay, Agilent is offering one of the few possibilities for selectively estimating miRNA before expression analysis. However, by now little is known about factors affecting miRNA analysis. Herein, the important impact of total RNA quality on quantification of mRNA and miRNA should be considered.
RNA quality control in miRNA expression analysis
Agilent Technologies Application Note 5990-5557EN
mRNA and microRNA quality control for RT-qPCR analysis
Methods. 2010 Apr;50(4): 237-243
The importance of high quality sample material, i.e. non-degraded or fragmented RNA, for classical gene expression profiling is well documented. Hence, the analysis of RNA quality is a valuable tool in the preparation of methods like RT-qPCR and microarray analysis. For verification of RNA integrity, today the use of automated capillary electrophoresis is state of the art. Following the recently published MIQE guidelines, these pre-PCR evaluations have to be clearly documented in scientific publication to increase experimental transparency. RNA quality control may also be integrated in the routine analysis of new applications like the investigation of microRNA (miRNA) expression, as there is little known yet about factors compromising the miRNA analysis. Agilent Technologies is offering a new lab-on-chip application for the 2100 Bioanalyzer making it possible to quantify miRNA in absolute amounts (pg) and as a percentage of small RNA (%). Recent results showed that this analysis method is strongly influenced by total RNA integrity. Ongoing RNA degradation is accompanied by the formation of small RNA fragments leading to an overestimation of miRNA amount on the chip. Total RNA integrity is known to affect the performance of RT-qPCR as well as the quantitative results in mRNA expression profiling. The actual study identified a comparable effect for miRNA gene expression profiling. Using a suitable normalization method could partly reduce the impairing effect of total RNA integrity.
and much more => http://RNA-integrity.gene-quantification.info
- Procedures for Quality Control of RNA Samples for Use in Quantitative Reverse Transcription PCR.
- Measurable impact of RNA quality on gene expression results from quantitative PCR.
- Preservation of fine-needle aspiration specimens for future use in RNA-based molecular testing.
- Maintaining RNA integrity in a homogeneous population of mammary epithelial cells isolated by Laser Capture Microdissection.
- RNA integrity in post-mortem samples: influencing parameters and implications on RT-qPCR assays.
- Analysis of Circulating MicroRNA - Pre-analytical and Analytical Challenges.
- Reliable gene expression measurements from degraded RNA by quantitative real-time PCR depend on short amplicons and a proper normalization.
- Evaluation of different RNA extraction methods for small quantities of plant tissue: Combined effects of reagent type and homogenization procedure on RNA quality-integrity and yield.
- Combined expression of miR-122a, miR-1, and miR-200b can differentiate degraded RNA samples from liver, pancreas, and stomach.
- Maintaining RNA integrity in a homogeneous population of mammary epithelial cells isolated by Laser Capture Microdissection.
- Robust microRNA stability in degraded RNA preparations from human tissue and cell samples.
- Assessment of mRNA and microRNA stabilization in peripheral Human Blood for Multicenter studies and Biobanks
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Did you miss the qPCR 2011 Symposium in Freising?
BioEPS is now presenting the qPCR 2011 eConference online via video stream!
With our new eConference streaming server a world-wide know-how transfer becomes possible in a fast and simple way!
qPCR 2011 Event now available NOW as eConfernce => http://eConference.qPCR2011.net
At the event around 500 attendee from 47 contries participated from 28th March - 1th April 2011. During the scientific symposium the presented talks were recorded in excellent sound quality and high movie resolution.
The qPCR 2011 eConference package contains 65 talks, grouped in following topics:
Single-Cell qPCR, Next Generation Sequencing, High throughput qPCR, MIQE guidelines, RNAi-miRNA, qPCR Data analysis, Circulating Nucleic Acids, Pre-analytical Steps, New qPCR Applications in Molecular Diagnostics, and more ... ...
qPCR 2011 Event now available NOW as eConfernce => http://eConference.qPCR2011.net
You can watch an in-depth demo presentation of our seminars and talks right here
=> http://eseminars.bioeps.com/player/?action=demo
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MIQE_qPCR APP for iPhone, iPad and iPod - iOS Universal
http://MIQE-press.gene-quantification.info
Get help from a special team of experts in qPCR while on the move. MIQE - qPCR helps you in reviewing scientific works and checking your own experiments, when qPCR is involved. Check your project's compliance to MIQE in minutes, have all required references to hand, and follow qPCR events and news.
http://itunes.apple.com/app/miqe-qpcr/id423650002?mt=8
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Forward Please send the qPCR NEWS to further scientists and friends who are interested in qPCR !
Best regards,
Michael W. Pfaffl
responsible Editor of the Gene Quantification Pages
http://www.gene-quantification.info
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