Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1-2
pubmed:dateCreated
2005-6-13
pubmed:abstractText
Despite the success and popularity of microarrays as a high-throughput technology for gene-expression studies, its sensitivity is as yet fairly limited. We have successfully combined the use of PCR-Select cDNA subtraction and Affymetrix GeneChips (AGC) to identify differentially expressed gene markers. Total RNA (totRNA) from combined hippocampus and cerebellum tissues of 2-week-old rat pups maintained for 5 weeks on an n-3 fatty acid (FA) deficient diet supplied to dams was isolated, SMART-amplified, and used for PCR-Select subtraction versus an adequately fed control litter preparation. Subtracted and amplified ds-cDNA end products were fragmented, terminally labeled with biotin-ddUTP and hybridized with RN-U34A AGC. At least 10-fold more potential gene markers with log2(T/D) > or = 1.4 were found versus the traditional AGC technology when the same chip was tested using nonsubtracted targets. Of this set of markers, 30% were robustly validated by real-time relative RT-PCR (rtrRT-PCR) and grouped as "confirmed" markers while the remaining were ascribed as "latent" markers. An improved and universal protocol to provide a rapid assessment for gene profiling in biological specimens is indicated.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jun
pubmed:issn
0169-328X
pubmed:author
pubmed:issnType
Print
pubmed:day
13
pubmed:volume
137
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
110-8
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:15950768-Animals, pubmed-meshheading:15950768-Animals, Newborn, pubmed-meshheading:15950768-Brain Chemistry, pubmed-meshheading:15950768-Cerebellum, pubmed-meshheading:15950768-DNA, Complementary, pubmed-meshheading:15950768-Fatty Acids, Omega-3, pubmed-meshheading:15950768-Food, Formulated, pubmed-meshheading:15950768-Gene Expression Profiling, pubmed-meshheading:15950768-Gene Expression Regulation, pubmed-meshheading:15950768-Genetic Markers, pubmed-meshheading:15950768-Hippocampus, pubmed-meshheading:15950768-Lipids, pubmed-meshheading:15950768-Nerve Tissue Proteins, pubmed-meshheading:15950768-Oligonucleotide Array Sequence Analysis, pubmed-meshheading:15950768-Polymerase Chain Reaction, pubmed-meshheading:15950768-RNA, Messenger, pubmed-meshheading:15950768-Rats, pubmed-meshheading:15950768-Rats, Sprague-Dawley, pubmed-meshheading:15950768-Stress, Physiological
pubmed:year
2005
pubmed:articleTitle
Improved representation of gene markers on microarray by PCR-Select subtracted cDNA targets.
pubmed:affiliation
Department of Neurobiology, Weizmann Institute of Science, Rehovot 76100, Israel.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't, Validation Studies