263 research outputs found

    Thresholds For Dielectric-Breakdown In Laser-Irradiated Diamond

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    We report on picosecond laser-induced damage experiments that were carried out on a natural type-IIa diamond and a thick specimen of high-quality chemically vapor-deposited (CVD) diamond. In conjunction with earlier measurements performed elsewhere on an \u27\u27optically thick\u27\u27 single crystal, it is shown that for spot sizes (2omega) ranging from 3 to 60 mum, the breakdown field strength (E(BD)) at the damage threshold of diamond obeys a pattern best described as follows: E(BD) congruent-to A/square-root 2omega, where A = 30.7 and 38.7 MV mu1/2/cm at 532 and 1064 nm, respectively. The case of CVD diamond demonstrates that if problems arising from localized high absorption at the deposition surface can be avoided, this material should be of much promise for contemplated high-power free-electron laser window applications

    Fully automated high-throughput chromatin immunoprecipitation for ChIP-seq: Identifying ChIP-quality p300 monoclonal antibodies

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    Chromatin immunoprecipitation coupled with DNA sequencing (ChIP-seq) is the major contemporary method for mapping in vivo protein-DNA interactions in the genome. It identifies sites of transcription factor, cofactor and RNA polymerase occupancy, as well as the distribution of histone marks. Consortia such as the ENCyclopedia Of DNA Elements (ENCODE) have produced large datasets using manual protocols. However, future measurements of hundreds of additional factors in many cell types and physiological states call for higher throughput and consistency afforded by automation. Such automation advances, when provided by multiuser facilities, could also improve the quality and efficiency of individual small-scale projects. The immunoprecipitation process has become rate-limiting, and is a source of substantial variability when performed manually. Here we report a fully automated robotic ChIP (R-ChIP) pipeline that allows up to 96 reactions. A second bottleneck is the dearth of renewable ChIP-validated immune reagents, which do not yet exist for most mammalian transcription factors. We used R-ChIP to screen new mouse monoclonal antibodies raised against p300, a histone acetylase, well-known as a marker of active enhancers, for which ChIP-competent monoclonal reagents have been lacking. We identified, validated for ChIP-seq, and made publicly available a monoclonal reagent called ENCITp300-1
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