Validating affymetrix chip results with real time pcr stark law backdating contracts
This technology has been used for gene expression, genotyping, mutation detection, and absolute quantization of nucleic-acid sequences  developed a high throughput SNP genotyping assay demonstrating high accuracy and call rate in human samples using a nanofluidic platform.
This new real-time PCR technology makes it possible to perform validation as well as high throughput gene expression measurements using very limited sample and reagent quantities .
Here we show that mi RNA expression profiling and validation are possible using high throughput real-time quantitative PCR (q PCR) method with the microfluidic technology.
FFPE and FF samples appear to perform similarly in this platform when c DNA are generated from 96 multiplexed RT reactions.
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.
Fourteen out of 16 mi RNA targets exhibited lower Ct values in the microfluidics 48.48 dynamic array system compared to those obtained by standard q PCR using the ABI 7900 HT (Figure ).
The mean Ct values between the platforms were 12.48 (± 0.49) for the 48.48 dynamic array and 16.21 (± 0.82) for the ABI 7900 HT (coefficient of variance CV = 0.08 and 0.06, respectively) reflecting a significantly increased sensitivity by the microfluidics array when q PCR reactions are being carried out in nanoliter volumes.
Eleven different mi RNA primers were used in single-plex (Y-axis) or 11-plex (X-axis) for reverse transcriptions using A549 (A) and H1299 (B) lung cancer cell lines.
Each plot displays mean values calculated from triplicate samples.
To evaluate the dynamic range of the multiplex RT-PCR in the 48.48 dynamic array systems, we evaluated the Ct values for each mi RNA target using total input RNA quantities at 10 ng, 25 ng, 50 ng or 100 ng per reaction.
Real-time quantitative PCR (q PCR) is considered a 'gold standard' for quantification of gene expression and has been widely employed as a validation method for microarray studies.
However, the q PCR method is a relatively low throughput, high cost, and tedious technique typically performed in a 96 or 384 well plate format.