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Adult fly versus embryo (0-16h)

ABSTRACT: Total RNA was isolated from wild-type adult flies (yellow white) and from embryos (cs, 0-16h) with TRIZOL reagent. 12 microgram of each total RNA were labeled with fluorescent dyes, following cDNA synthesis with amino-allyl dUTP in addition to the four natural dNTPs using a 1:1 mixture of oligo(dT) and random nonamer primers. An all pairs experiment design type is where all labeled extracts are compared to every other labeled extract. Keywords: all_pairs

ORGANISM(S): Drosophila melanogaster

PROVIDER: GSE3581 | GEO | 2006/02/28

SECONDARY ACCESSION(S): PRJNA93667

REPOSITORIES: GEO

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Adult fly versus embryo (0-16h)

Project description:Total RNA was isolated from wild-type adult flies (yellow white) and from embryos (cs, 0-16h) with TRIZOL reagent. 12 microgram of each total RNA were labeled with fluorescent dyes, following cDNA synthesis with amino-allyl dUTP in addition to the four natural dNTPs using a 1:1 mixture of oligo(dT) and random nonamer primers. An all pairs experiment design type is where all labeled extracts are compared to every other labeled extract. Keywords: all_pairs Computed

2010-07-01 | E-GEOD-3581 | biostudies-arrayexpress

Transcription profiling of adult fly vs embryo (0-16h), Gerber AP. 2005

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Wild-type versus Pumilio mutant flies

Project description:Total RNA was isolated with TRIZOL reagent from wild-type flies (yellow white, yw) and Pumilio mutant flies (pum13; Barker DD. et al. (1992) Genes Dev 6, 2312-26). 12 microgram of total RNA was used for cDNA synthesis using a 1:1 mixture of oligo(dT) and random nonamer primers and in the presence of amino-allyl dUTP. Set of arrays that are part of repeated experiments Keywords: Biological Replicate

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Khd1p associated mRNAs

Project description:RNA-coimmunopurifications with TAP-tagged Khd1 protein from Saccharomyces cereviseae. Untagged strain (BY4741) served as a control (Gerber et al., 2004). Cells were grown to midlog phase in rich media and harvested by centrifugation. TAP-tagged Khd1 was affinity purified from cell-free extracts with IgG sepharose and eluted with TEV protease. RNA was isolated from extract (=input) and from purified protein samples by phenol-chloroform extraction. RNAs were reverse transcribed using a mixture of oligo-dT and random nonamer oligos in the presence of amino-allyl dUTP/ dNTP mixture. cDNAs were fluorescently labeled and hybridized on yeast DNA microarrays over night at 65 degrees. For a detailed procedure see http://microarray-pubs.stanford.edu/yeast_puf and also Gerber AP et al. PLoS Biology, 2004. An all pairs experiment design type is where all labeled extracts are compared to every other labeled extract. Strain Name: yeast strain with/without TAP-tagged Khd1 Keywords: all_pairs

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Wild-type versus Pumilio mutant flies

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Puf3delta vs. wild type comparison

Project description:Three independent experiments: S. cervisiae wild-type (BY4741) and Puf3 mutant cells were grown in minimal medium supplemented with 3% glycerol. Cells were harvested in mid-log phase by centrifugation and total RNA was prepared by hot-phenol extraction. cDNA was prepared with oligo-dT and using a mixture of amino-allyl dUTP and dNTPs, fluorescently labeled (Cy3= wild-type, Cy5 = mutant) and hybridize on S. cerevisiae DNA microarray. Set of arrays organized by shared biological context, such as organism, tumors types, processes, etc. Keywords: Logical Set

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Slf1p and Sro9p affinity isolations

Project description:To identify RNAs specifically associated with Slf1p and Sro9p, cells expressing TAP-tagged Slf1, Sro9 or Fpr1 (negative control) were grown to mid-log phase in synthetic complete medium and harvested by centrifugation. RNA affinity isolations were essentially performed as described (Gerber et al. 2004 PLoS Biol.). In brief, TAP-tagged protein were captured from cell extracts with IgG coupled agarose beads (Sigma) and released by incubation with a site-specific protease (AcTEV, Sigma). from the extract (input) and from the affinity isolates was purified with the RNeasy Mini/ Micro Kit (Qiagen). RNAs were reverse transcribed using a mixture of oligo-dT and random nonamer oligos in the presence of amino-allyl dUTP/dNTP mixture. cDNAs were fluorescently labeled and hybridized on yeast oligo microarrays over night at 42 degrees in formamide-based hybridization buffer.

2012-02-08 | GSE27310 | GEO

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