The T7-primer is a source of experimental bias and introduces variability between microarray platforms.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 18431470.
- Also identified by DOI 10.1371/journal.pone.0001980 and PMC identifier 2292241.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
Abstract
Eberwine(-like) amplification of mRNA adds distinct 6-10 bp nucleotide stretches to the 5' end of amplified RNA transcripts. Analysis of over six thousand microarrays reveals that probes containing motifs complementary to these stretches are associated with aberrantly high signals up to a hundred fold the signal observed in unaffected probes. This is not observed when total RNA is used as target source. Different T7 primer sequences are used in different laboratories and platforms and consequently different T7 primer bias is observed in different datasets. This will hamper efforts to compare data sets across platforms.
Medical subject headings
- Bacteriophage T7
- DNA Primers
- Oligonucleotide Array Sequence Analysis