Validating affymetrix chip results with real time pcr
Five years later, what was so recently a brand-new technology has become a laboratory mainstay.
Frost & Sullivan, San Antonio, Texas, which analyzes emerging high-tech and industrial markets, projects a world microarray market of over 0 million this year, and predicts that it will be closing in on a billion dollars annually by 2010.
Furthermore, archival formalin fixed, paraffin embedded (FFPE) samples are increasingly becoming the primary resource for gene expression studies because fresh frozen (FF) samples are often difficult to obtain and requires special storage conditions.
In this study, we evaluated the mi RNA expression levels in FFPE and FF samples as well as several lung cancer cell lines employing a high throughput q PCR-based microfluidic technology.
The most traditional method for characterizing m RNA transcripts is northern blot analysis.
We compared the accuracy of microarray measurements obtained with oligonucleotide arrays (Gene Chip, Affymetrix) with a laboratory-developed c DNA array by assaying test RNA samples from an experiment using a paradigm known to regulate many genes measured on both arrays.
Although determination of relative quantity is widely accepted as a reliable means of measuring differences between samples, there are advantages to being able to determine the absolute copy numbers of a given target.
One approach to absolute quantification relies on construction of an accurate standard curve using appropriate external standards of known concentration.
But the journey from a chip to a target to a phase III trial faces some major roadblocks.Quantitative PCR is a rapid and highly sensitive technique for accurate quantification of microarray results; however, careful consideration of experimental design, quality of primer/probe design, internal standards, and normalization procedures are pivotal, particularly when the work involves postmortem tissue.In the late 1990s, you could measure the market in DNA microarrays in the tens of millions of dollars.Micro RNAs (mi RNAs) represent a growing class of small non-coding RNAs that are important regulators of gene expression in both plants and animals.Studies have shown that mi RNAs play a critical role in human cancer and they can influence the level of cell proliferation and apoptosis by modulating gene expression.