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Applied Biosystems Unveils TaqMan® PreAmp Master Mix Kit

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Applied Biosystems, an Applera Corporation business, has announced the launch of a commercial reagent mix that is designed to enable quantitative, real-time gene expression analysis on very small or rare samples.

The TaqMan® PreAmp Master Mix Kit is intended to address the challenge faced by researchers working with precious samples to extract enough mRNA to perform advanced gene expression analyses using real-time PCR.

Previous commercially available pre-amplification methods designed to analyze small samples using real-time PCR have been limited in their effectiveness because they inconsistently amplify the starting material, resulting in biased and inaccurate gene amplification.

These methods are also typically laborious and time-intensive for researchers, with some methods requiring ten hours of manual steps and PCR cycling time.

As a result, many of these samples remain archived as opposed to being studied for the important molecular information they may contain.

The TaqMan PreAmp Master Mix Kit is designed to address this challenge with an optimized formulation that works with a multiplexed pool of up to 100 TaqMan Gene Expression Assays.

The result is a 1,000-16,000-fold increase in gene transcripts available for analysis.

The simple process is designed to enable researchers to perform uniform and unbiased amplification of as little as one nanogram of cDNA, and takes only 15 additional minutes of hands-on time and 1.5 hours of PCR cycling time.

Researchers can then conduct up to 200 real-time PCR reactions per pre-amplification reaction without compromising their available sample material.

Researchers at the Fox Chase Cancer Center are using the TaqMan PreAmp Master Mix Kit to analyze its laser microdissected samples of hepatitis C-infected liver cells.

"Using the TaqMan PreAmp Master Mix Kit we achieved nearly 100% efficient amplification of our target sequences with reproducible results using varying numbers of cells as starting material," said Renata Coudry, M.D., Ph.D., manager for Fox Chase Cancer Center's laser capture microdissection facility.

"The balance of targets in the sample remains the same, enabling truly quantitative analysis."

"This new amplification method will expand the use of gene expression profiling to characterize limited cell populations, including virus-infected cells."

The TaqMan PreAmp Master Mix Kit accepts from one to 250 nanograms of cDNA generated using random primers.

The kit is optimized for use with the TaqMan Gene Expression Assay product line and the Applied Biosystems family of Real-Time PCR Systems.

"We developed the TaqMan PreAmp Master Mix Kit in response to requests from clinical researchers working with laser capture microdissected samples, needle biopsies, paraffin-embedded tissues and other samples with limited amounts of RNA," said Carl Hull, vice president and general manager for sequence detection systems at Applied Biosystems.

"As a result, our customers now have access to a robust way to stretch limited samples, perform real-time, quantitative analysis on more gene targets and even conduct gene expression analysis on samples that were previously impossible to study."