Project description:This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/ Teladorsagia circumcincta, the brown stomach worm, is the most important gastro-intestinal parasite of sheep in temperate regions and is present on 100% of sheep farms in the UK. Infection with T. circumcincta causes a spectrum of disease in the host, ranging from weight loss to death, and results in significant production losses. Parasitic disease is currently controlled by treatment with anthelmintics but resistance to these drugs is widespread and threatens the viability of the livestock industry. There are now well-documented cases of UK farms harbouring triple-resistant T. circumcincta, which can no longer be controlled by any of the three major anthelmintic classes, forcing farmers to abandon sheep rearing. High-throughput sequencing of life stages of T. circumcincta will be used for gene finding and transcriptome analysis.
Project description:To interrogate the glycoproteome following knockdown of pglL and subsequent knockdown of glycosylation in Burkholderia cenocepacia through Zwitterionic Hydrophilic Interaction Liquid Chromatography (ZIC-HILIC) glycopeptide enrichment with 0.1% rhamnose for dCas9 induction. Using strains harbouring the pglL-targeting guide C (Bc-pglL-C) and guideless plasmids.
Project description:To interrogate the glycoproteome following knockdown of pglL and subsequent knockdown of glycosylation in Burkholderia cenocepacia through Zwitterionic Hydrophilic Interaction Liquid Chromatography (ZIC-HILIC) glycopeptide enrichment with 0.1% rhamnose for dCas9 induction. Using strains harbouring the pglL-targeting guide C (Bc-pglL-C) and guideless plasmids.
Project description:To interrogate the glycoproteome following knockdown of pglL and subsequent knockdown of glycosylation in Burkholderia cenocepacia through Zwitterionic Hydrophilic Interaction Liquid Chromatography (ZIC-HILIC) glycopeptide enrichment with 0.1% rhamnose for dCas9 induction– loading control pre-enrichment. Using strains harbouring the pglL-targeting guide C (Bc-pglL-C) and guideless plasmids.
Project description:Haploid androgenetic stem cells (haSCs) are invaluable resources for studying animal traits and greatly impact livestock breeding, but livestock haSCs have yet to be obtained. Here, we report the derivation of cattle and sheep haSCs using the recombined FACE medium, and these cells harbor formative pluripotency features. Notably, knockout cattle and sheep can be obtained upon injection of the protaminized haSCs with targeted gene mutations into intact oocytes.
Project description:To test whether the addition of a peptide nucleic acid (PNA) clamp, which binds WT KRAS at codon 12, can increase the efficacy of mutation detection for KRASG12D within a targeted NGS setting. We tested the effect of clamping the wild-type KRAS sequence in a reference standard (Tru-Q 7, 1.3% Tier from Horizon Diagnostics, Cambridge, UK) with a KRAS c.35G>A mutation (KRASG12D) at an allelic frequency (AF) of 1.3% assessed by digital droplet PCR (ddPCR). We then re-tested the PNA on circulating-free DNA from a patient harbouring a KRASG12D mutation (at an AF of 3.2%, determined by ddPCR). Multiple runs were conducted using 10, 5, 2.5 and 1ng of DNA input.
