ABSTRACT: Use of the Ion Torrent PGM for Determining the Genomic Sequences of Francisella and Coxiella-Like Endosymbionts and Rickettsia Directly from Hard Ticks
Project description:Use of the Ion Torrent PGM for Determining the Genomic Sequences of Francisella and Coxiella-Like Endosymbionts and Rickettsia Directly from Hard Ticks
Project description:Use of the Ion Torrent PGM for Determining the Genomic Sequences of Francisella and Coxiella-Like Endosymbionts and Rickettsia Directly from Hard Ticks
Project description:Use of the Ion Torrent PGM for Determining the Genomic Sequences of Francisella and Coxiella-Like Endosymbionts and Rickettsia Directly from Hard Ticks
Project description:Use of the Ion Torrent PGM for Determining the Genomic Sequences of Francisella and Coxiella-Like Endosymbionts and Rickettsia Directly from Hard Ticks
Project description:Use of the Ion Torrent PGM for Determining the Genomic Sequences of Francisella and Coxiella-Like Endosymbionts and Rickettsia Directly from Hard Ticks
Project description:Next generation sequencing using the Ion Torrent PGM platform after PNLDC1 silencing in mouse embryonic stem cells E14 using esiRNA
Project description:The ligation step in RNA sequencing library generation is a known source of bias. We present the first comparison of the standard duplex adaptor protocol supplied by Life Technologies for use on the Ion Torrent PGM with an alternate single adaptor approach involving CircLigase (CircLig). We also investigate whether using the thermostable ligase Methanobacterium thermoautotrophicum RNA ligase K97A (Mth K97A) for the initial ligation step in the CircLigase protocol reduces bias. A pool of small RNA fragments of known composition was converted into a sequencing library using one of three protocols and sequenced on an Ion Torrent PGM. The single adaptor CircLigase-based approach significantly reduces, but does not eliminate, bias in Ion Torrent data. Using Mth K97A as part of the CircLig method does not further reduce bias.
Project description:Ticks (Acari: Ixodida) feed exclusively on blood which lacks essential nutrients such as vitamins and cofactors. These deficiencies are presumably complemented through specific symbiotic microorganisms such as Coxiella – Like Endosymbionts (CLEs) of Rhipicephalus ticks. CLE are localized in specialized host tissue cells within the Malpighian tubules (Mt) and the ovaries (Ov) from which they are maternally transmitted to developing oocytes. These two organs differ in function and cell types, and possibly also in activities performed by CLE occupants. To get insights into CLE functions within these two organs, we used comparative proteomics of CLEs in Mt and Ov of unfed R. sanguineus ticks
Project description:Exosomes/microvesicles (hereafter referred to as extracellular vesicles) were isolated from the ULF of day 14 cyclic and pregnant ewes using ExoQuick-TC. Extracellular vesicle RNA was pooled (n=4 per status) and analyzed for small RNAs by sequencing on the Ion Torrent PGM platform and analysis with CLC Genomics Workbench small RNA workflow based on the miRBase (Release 19) Bos taurus database.