Natural genetic variation in yeast longevity
Ontology highlight
ABSTRACT:
ORGANISM(S): Saccharomyces cerevisiae
SUBMITTER: jianliang dai
PROVIDER: E-GEOD-37590 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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2007-04-16 | E-TABM-112 | biostudies-arrayexpress
Project description:We measured mRNA abundance in the seedling leaves of 35 recombinant Steptoe x Morex doubled-haploid lines (no replicates) and in parental genotypes, Steptoe and Morex, 3 replicates each, total 41 chip.
2007-04-11 | E-TABM-111 | biostudies-arrayexpress
Project description:The goal of this study was to investigate the generation and normalization of microRNA expression data based on microarray technologies to comparatively assess their quality. Two profiling platforms were compared: the single-channel Affymetrix GeneChip(r) and the Exiqon dual-channel miRCURY LNA(tm), which was processed as a single-channel array. Due to fundamental differences in the platform constitution, the normalization methods developed for gene expression need to be applied very cautiously to microRNA raw data. This motivated the development of a novel normalization method based on controllable assumptions, which uses the intensities of spike-in control probes. The results showed that the novel normalization method reduced the data variability in the most consistent way and confirmed the reliability of the differentially expressed microRNAs obtained, based on an RT-qPCR experiment performed for a subset of microRNAs. The conclusion was that the Exiqon platform combined with the novel spike-in controls based normalization method provides high-quality microRNA expression data suitable for reliable downstream analysis. This preprocessing pipeline was implemented into an R package called ExiMiR and deposited in the Bioconductor repository. The Exiqon data set has also been deposited in ArrayExpress under accession number E-MTAB-876.
2012-04-01 | E-MTAB-875 | biostudies-arrayexpress
Project description:The stability, localization and translation of mammalian mRNAs are largely determined by sequences in the 3' untranslated region (UTR). Here, we describe a conserved program of increased upstream polyadenylation site usage following activation of primary murine CD4+ T lymphocytes. This program, resulting in shorter 3' UTRs, is a characteristic of immune cell activation and strongly correlates with proliferation across diverse cell types and tissues. Enforced expression of full-length 3' UTRs confers differences in protein expression that can in some cases be eliminated by deletion of predicted microRNA target sites in the variably included region. Together, our data indicate that polyadenylation site usage is coordinately regulated such that states of increased proliferation are associated with widespread reduction of 3' UTR-based regulation. Keywords: activation, exon array, time course Two or three biological replicates per time point
2010-05-26 | E-GEOD-10666 | biostudies-arrayexpress
Project description:Goal: To identify copy number variation in normal individuals using high density, non-polymorphic oligonucleotide probes Background DNA sequence diversity within the human genome may be more greatly affected by copy number variations (CNVs) than single nucleotide polymorphisms (SNPs). Although the importance of CNVs in genome wide association studies (GWAS) is becoming widely accepted, the optimal methods for identifying these variants are still under evaluation. We have previously reported a comprehensive view of CNVs in the HapMap DNA collection using high density 500K EA (Early Access) SNP genotyping arrays which revealed greater than 1,000 CNVs ranging in size from 1kb to over 3Mb. Although the arrays used most commonly for GWAS predominantly interrogate SNPs, CNV identification and detection does not necessarily require the use of DNA probes centered on polymorphic nucleotides and may even be hindered by the dependence on a successful SNP genotyping assay. Results In this study, we have designed and evaluated a high density array predicated on the use of non-polymorphic oligonucleotide probes for CNV detection. This approach effectively uncouples copy number detection from SNP genotyping and thus has the potential to significantly improve probe coverage for genome-wide CNV identification. This array, in conjunction with PCR-based, complexity-reduced DNA target, queries over 1.3M independent NspI restriction enzyme fragments in the 200bp to 1100bp size range, which is a several fold increase in marker density as compared to the 500K EA array. In addition, a novel algorithm was developed and validated to extract CNV regions and boundaries. Conclusions Using a well-characterized pair of DNA samples, close to 200 CNVs were identified, of which nearly 50% appear novel yet were independently validated using quantitative PCR. The results indicate that non-polymorphic probes provide a robust approach for CNV identification, and the increasing precision of CNV boundary delineation should allow a more complete analysis of their genomic organization. A set of five genomic DNA samples containing different numbers of X chromosomes (1X to 5X sample set, including NA15510 and NA10851) were hybridized to Nsp copy number (CN) arrays in triplicate to evaluate detection of copy number variation using high density, non-polymorphic oligonucleotide probes. 6 Hapmap samples were hybridized to Nsp CN arrays to evaluate Mendelian inheritance of CNVs.
2008-04-24 | E-GEOD-9053 | biostudies-arrayexpress
Project description:To map meiotic crossovers and noncrossovers across the yeast genome, we genotyped the four spores of 51 tetrads arising from sporulation of a YJM789/S96 hybrid strain. Sporulation was induced, tetrads were dissected, and genomic DNA from each spore was hybridized to microarrays, one array per spore. 12 YJM789 and 13 S96 hybridizations were also performed as a reference for genotyping. To analyze recombination mutants, we also hybridized DNA from 5 tetrads arising from a YJM789/S96 hybrid strain homozygous for a MSH4 deletion, and from 20 spores (6 dyads and 8 single spores) arising from a hybrid homozygous for a MMS4 deletion.
2009-08-04 | E-TABM-470 | biostudies-arrayexpress
Project description:Aneuploidy has been well documented in blastocyst embryos, but prior studies have been limited in scale and/or lack mechanistic data. We previously reported preclinical validation of microarray 24-chromosome preimplantation genetic screening (PGS) in a 24-hour protocol. The method diagnoses chromosome copy number, structural chromosome aberrations, parental source of aneuploidy, and distinguishes certain meiotic from mitotic errors. In this study our objective was to examine aneuploidy in human blastocysts and determine correspondence of karyotypes between trophectoderm (TE) and inner cell mass (ICM). We disaggregated 51 blastocysts from seventeen couples into ICM and one or two TE fractions. The average maternal age was 31. Next, we ran 24-chromosome microarray molecular karyotyping on all of the samples, and then performed a retrospective analysis of the data. The average per-chromosome confidence was 99.95%. Approximately 80% of blastocysts were euploid. The majority of aneuploid embryos were simple aneuploid, i.e., one or two whole-chromosome imbalances. Structural chromosome aberrations, which are common in cleavage stage embryos, occurred in only three blastocysts (5.8%). All TE biopsies derived from the same embryos were concordant. Forty-nine of fifty-one (96.1%) inner cell mass (ICM) samples were concordant with TE biopsies derived from the same embryos. Discordance between TE and ICM occurred only in the two embryos with structural chromosome aberration. We conclude that trophectoderm karyotype is an excellent predictor of inner cell mass karyotype. Discordance between TE and ICM occurred only in embryos with structural chromosome aberrations. We thawed all fractions at 22°C, and then added Arcturus PicoPure Lysis Buffer (Molecular Devices, Sunnyvale, CA) to each of the biopsies. The tubes were incubated at 56°C for one hour, and then heat inactivated at 95°C for 10 minutes. DNA from the lysed biospsies was amplified using a commercial kit (GE Healthcare, Waukesha, WI) for multiple displacement amplification (MDA). MDA reactions were incubated at 30°C for 2.5 hours and then heat-inactivated at 65°C for five minutes. The amplified samples were genotyped using Illumina (San Diego, CA, USA) Infinium II genotyping microarrays (CytoSNP-12 chips) using a modified 24-hour protocol, as described previously*. Previously, we developed a genotyping microarray molecular karyotyping technology that uses parental genetic data to increase accuracy and determine mechanism and source of aneuploidy*. The algorithm uses parental genotypes and the observed distribution of unprocessed single cell microarray channel intensities to diagnose whole-chromosome imbalances and structural chromosome aberrations*. Because parental genotypes are available, the algorithm readily identifies parental source of whole-chromosome imbalances and structural chromosome aberrations. Additionally, the algorithm uses parental information, high confidence disomic single cell measurements on children, and recombination probabilities (genome.ucsc.edu) to determine the phase of the parental chromosomes. The phased data is then used to determine whether certain trisomies and uniparental disomies were mitotic or meiotic in origin*. *Johnson DS, Gemelos G, Baner J, Ryan A, Cinnioglu C, Banjevic M, Ross R, Alper M, Barrett B, Frederick J, Potter D, Behr B, Rabinowitz M. (2010). Preclinical validation of a microarray method for full molecular karyotyping of blastomeres in a 24-hour protocol. Human Reproduction January 24 [Epub ahead of print].
2010-09-28 | E-GEOD-22864 | biostudies-arrayexpress
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