A New Approach to Gene Expression Analysis

Most of the attention on gene expression analysis and discovery tools has centered on microarray technology, but alternative methods, including SAGE (serial analysis of gene expression)1 and direct cDNA sequencing, do exist. Another such technique is Hayward, Calif.-based Lynx Therapeutics' Massively Parallel Signature Sequencing (MPSS™) technology, a method for counting millions of mRNA molecules from a given sample. Like SAGE, MPSS quantifies tagged cDNA fragments; but whereas SAGE uses

Written byAileen Constans
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Norman Russell, Lynx president and CEO, compares gene expression analysis to a puzzle, noting that a complete picture is not possible unless every piece is present: "From a systems biology standpoint, what you need to do is to start with a jigsaw puzzle with all the pieces," he says. With that in mind, the MPSS process begins by using Lynx's Megaclone™ technology to create a library, in which every mRNA molecule in a sample is represented by a single 5-µm microbead. First, amplified cDNAs prepared from a sample are digested with DpnII. The 3'-most DpnII-poly-(A) fragments are then selected, tagged with one of more than 16 million different 32-base oligonucleotides, hybridized, and ligated to microbeads containing oligonucleotides complementary to the original tags. Each microbead has on its surface thousands of copies of one of the 32-base tags, ensuring that all amplified cDNA fragments in the sample that contain the associated ...

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