Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Mismatch oligonucleotides in human and yeast


ABSTRACT: Background Mismatched oligonucleotides are widely used on microarrays to differentiate specific from nonspecific hybridization. While many experiments rely on such oligos, the hybridization behavior of various degrees of mismatch (MM) structure has not been extensively studied. Here, we present the results of two large-scale microarray experiments on S.cerevisiae and H.sapiens genomic DNA, to explore MM oligonucleotide behavior with real sample mixtures under tiling-array conditions. Results We examined all possible nucleotide substitutions at the central position of 36-nucleotide probes, and found that nonspecific binding by MM oligos depends upon the individual nucleotide substitutions they incorporate: C->A, C->G and T->A (yielding purine-purine mispairs) are most disruptive, whereas A->X were least disruptive. We also quantify a marked GC skew effect: substitutions raising probe GC content exhibit higher intensity (and vice versa). This skew is small in highly-expressed regions (±0.5% of total intensity range) and large (±2% or more) elsewhere. Multiple mismatches per oligo are largely additive in effect: each MM added in a distributed fashion causes an additional 21% intensity drop relative to PM, three-fold more disruptive than adding adjacent mispairs (7% drop per MM). This SuperSeries is composed of the following subset Series: GSE13172: Mismatch oligonucleotides in human GSE13174: Mismatch oligonucleotides in yeast Refer to individual Series

ORGANISM(S): Homo sapiens

SUBMITTER: Michael Seringhaus 

PROVIDER: E-GEOD-13175 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Mismatch oligonucleotides in human and yeast: guidelines for probe design on tiling microarrays.

Seringhaus Michael M   Rozowsky Joel J   Royce Thomas T   Nagalakshmi Ugrappa U   Jee Justin J   Snyder Michael M   Gerstein Mark M  

BMC genomics 20081231


<h4>Background</h4>Mismatched oligonucleotides are widely used on microarrays to differentiate specific from nonspecific hybridization. While many experiments rely on such oligos, the hybridization behavior of various degrees of mismatch (MM) structure has not been extensively studied. Here, we present the results of two large-scale microarray experiments on S. cerevisiae and H. sapiens genomic DNA, to explore MM oligonucleotide behavior with real sample mixtures under tiling-array conditions.<h  ...[more]

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