Increased DNA microarray hybridization specificity using sscDNA targets.

ABSTRACT:

Background

The most widely used amplification method for microarray analysis of gene expression uses T7 RNA polymerase-driven in vitro transcription (IVT) to produce complementary RNA (cRNA) that can be hybridized to arrays. However, multiple rounds of amplification are required when assaying very small amounts of starting RNA. Moreover, certain cRNA-DNA mismatches are more stable than the analogous cDNA-DNA mismatches and this might increase non-specific hybridization. We sought to determine whether a recently developed linear isothermal amplification method (ribo-SPIA) that produces single stranded cDNA would offer advantages over traditional IVT-based methods for microarray-based analyses of transcript expression.

Results

A single round of ribo-SPIA amplification produced

SUBMITTER: Barker CS 

PROVIDER: S-EPMC1090574 | biostudies-literature | 2005 Apr

REPOSITORIES: biostudies-literature