ChIP-seq of HA-tagged proteins in fungi

Strains were grown in the indicated conditions and cross-linking was performed as described previously[42 (link)]. Press-dried mycelia was frozen in liquid nitrogen and stored at -80˚C until chromatin preparation, which was performed as described previously[43 (link)]. For immunoprecipitation, 2 ug of anti-HA antibody (F7, Santa Cruz) was used. Library preparation was carried out according to[44 (link)] with the exception of the first end-repair step, which was replaced by the NEBNext End-Repair module (Cat. no. E6050). Libraries were checked and concentration determined using DNA High Sensitivity Bioanalyzer assay, mixed in equal molar ratio as described[44 (link)] and subjected to sequencing using Illumina HiSeq2500.

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Ries L.N., Pardeshi L., Dong Z., Tan K., Steenwyk J.L., Colabardini A.C., Ferreira Filho J.A., de Castro P.A., Silva L.P., Preite N.W., Almeida F., de Assis L.J., dos Santos R.A., Bowyer P., Bromley M., Owens R.A., Doyle S., Demasi M., Hernández D.C., Netto L.E., Pupo M.T., Rokas A., Loures F.V., Wong K.H, & Goldman G.H. (2020). The Aspergillus fumigatus transcription factor RglT is important for gliotoxin biosynthesis and self-protection, and virulence. PLoS Pathogens, 16(7), e1008645.

Publication 2020

anti-HA antibody (F7, HiSeq2500

Anti antibody Assay Chromatin Frozen Immunoprecipitation Library Molar Mycelia Nitrogen Sensitivity Strains

Corresponding Organization : Instituto Butantan

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Variable analysis

independent variables

Strains were grown in the indicated conditions

dependent variables

Chromatin preparation
Immunoprecipitation using 2 ug of anti-HA antibody (F7, Santa Cruz)
Library preparation and sequencing using Illumina HiSeq2500

control variables

Cross-linking was performed as described previously[42]
Chromatin preparation was performed as described previously[43]
Library preparation was carried out according to[44] with the exception of the first end-repair step, which was replaced by the NEBNext End-Repair module (Cat. no. E6050)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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