Project description:Investigation of whole genome gene expression level changes in F. verticillioides FRC M-3125 when exposed to 50 ug/ml 2-benzoxazolinone (BOA). Cultures were harvested two hours after exposure. Assessed in reference to control cultures of M-3125 exposed to ethanol (1% final concentration) since BOA was dissolved in ethanol.
Project description:Investigation of whole genome gene expression level changes in F. verticillioides FRC M-3125 when exposed to 50 ug/ml 2-benzoxazolinone (BOA). Cultures were harvested two hours after exposure. Assessed in reference to control cultures of M-3125 exposed to ethanol (1% final concentration) since BOA was dissolved in ethanol. A four chip study using total RNA recovered from two separate control cultures of F. verticillioides FRC M-3125 in PDB containing 1% ethanol and two separate experimental cultures of M-3125 grown in PDB plus the added treatment of 50 ug/ml ethanolic BOA. The cultures were incubated for two hours on a rotary shaker (200 rpm) at 27C in the dark and then harvested by vacuum filtration and frozen in liquid nitrogen. The chips were a F. verticillioides custom 14K array based on platform GPL14669.
Project description:The utility of RADseq in an experimental setting is also demonstrated, based on our chasacterisation of an APOBEC mutation signature in an APOBEC3A transfected mouse cell line. 0D5 cells, derived from SSM3 cells, were co-transfected with a mixture containing pcDNA3.1 vectors expressing either APOBEC3A or APOBEC3B (kindly donated by Vincent Caval), pcDNA3.1 construct expressing deaminase null APOBEC3A linked to a uracil deglycosylase construct and a plasmid encoding mutant GFP and WT mCherry that is a reporter for APOBEC mutagenesis. Cells were grown, and gDNA extracted, prior to preparation of RADseq libraries using a PstI- MspI double-digest. Libraries underwent a Pippin Prep to select fragments in the size range of 220-520 bp (genomic sequence plus 148 bp of adapters). Single-end sequencing (1x101bp) was performed on an Illumina NovaSeq6000 utilizing v1.5 chemistry. Reads were aligned to mm10 using bwa mem and variants called using the GATK4 pipeline.