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4abd8d2a-485c-4457-a98a-15239ce7cc97 - samples

ABSTRACT: This dataset contains counts for 699 tumor samples profiled by RNA-seq for the entire transcriptome for samples originating from OAK (GO28915).

PROVIDER: EGAD00001008628 | EGA |

REPOSITORIES: EGA

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A comprehensive RNA handling and transcriptomics guide for high-throughput processing of Plasmodium blood-stage samples.

Project description:Here we have designed a full processing pipeline of P. falciparum RNA that innclues RNA preservation, extraction, cDNA synthesis and amplification, globin depletion and final transcriptome readout. To validate the protocol we have analyzed transcriptomes of P. falciaprum samples derived from filed studies as well as 3D7 lab strains. This dataset have been used to establish the best conditions for whole transcriptome amplification protocol. Additionally it inclues the validation of bead extraction protocol. It also contains field samples used for validation of the entire protocol.

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Project description:Expression diversity of P. ramorum isolates belonging to the NA1 clonal lineage growing on solid CV8 was examined. It was found that although all the analyzed isolates belonged to a single clonal lineage, expression patterns were distinctive between isolates originating from coast live oak and California bay laurel. Global expression patterns of 13 isolates originating from coastal live oak and California bay laurel was investigated. No biological replicates were generated. The sequenced strain Pr102 was included. Gene models Phytophthora ramorum v1.0 were used to construct NimbleGen 72K x4 custom arrays.

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Project description:Tree ring features are affected by environmental factors and therefore are the basis for dendrochronological studies to reconstruct past environmental conditions. Oak wood often provides the data for these studies because of the durability particularly of oak heartwood and, hence the availability of samples spanning long time periods of the distant past. Wood formation is regulated in part by epigenetic mechanisms such as DNA methylation. Studies in the methylation state of DNA preserved in oak heartwood thus could identify epigenetic tree ring features informing on past environmental conditions. We investigated the feasibility of such studies using heartwood samples core-drilled from the trunks of standing oak trees spanning the AD 1776-2014. Heartwood contains little DNA, and large amounts of phenolic compounds known to hinder the preparation of high-throughput sequencing libraries. We sequenced whole-genome and DNA methylome libraries for oak heartwood up to 100 and 50 years of age, respectively. However, only 56 genomic regions with sufficient coverage for quantitative methylation analysis were identified, suggesting that the high-throughput sequencing of DNA will be in principal feasible for wood formed <100 years ago is impeded by the reduction in library complexity caused by the bisulfite treatment used to generate the oak methylome.

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