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SRX21046507: GSM7622269: P23-45_20min_input_gp96_replica1; Thermus thermophilus HB8; ChIP-Seq
1 ILLUMINA (Illumina HiSeq 4000) run: 2.1M spots, 313.4M bases, 125.3Mb downloads

External Id: GSM7622269_r1
Submitted by: Laboratory of Gene Technology, Biosystems, KU Leuven
Study: Chromatin immunoprecipitation DNA-sequencing (ChIP-seq) for RNA-polymerases encoded by P23-45 bacteriophage
show Abstracthide Abstract
By doing ChIP-seq with antibodies against two P23-45 RNA polymerases gp64 and gp96 we showed that both RNA polymerases interact with the phage genome at the early stage of infection. Gp96 interacts with pre-early P23-45 genes, while gp64 interacts with pre-early, early and some middle-stage genes. Overall design: Thermus thermophilus HB8 cells were infected with P23-45 bacteriophage at a high multiplicity of infection. Next, cells were collected at 5, 20, and 40 minutes post infection with the phage. Chromatin immunoprecipitation followed by DNA-sequencing (ChIP-seq) was performed using antibodies against P23-45 RNA polymerases gp64 and gp96 (first 1540 amino acid residues). Samples without addition of antibodies were used as controls for input DNA.
Sample: P23-45_20min_input_gp96_replica1
SAMN36493581 • SRS18317356 • All experiments • All runs
Library:
Name: GSM7622269
Instrument: Illumina HiSeq 4000
Strategy: ChIP-Seq
Source: GENOMIC
Selection: ChIP
Layout: PAIRED
Construction protocol: Infected cells were fixed with 1% formaldehyde for 20 minutes. The cross-linking reaction was stopped by the addition of dry glycine up to the final concentration of 0.5 M. Cells were incubated for 20 min with agitation. The cells were harvested by centrifugation and washed twice with ice-cold TBS (10 mM Tris-HCl pH 7.6, 150 mM NaCl). The cells were lysed and DNA was sheared by sonication to an average size of 200 – 500 bp. Immunoprecipitation was performed using corresponding antibodies and Sera-Mag SpeedBeads Protein A/G Sepharose magnetic beads (GEHealth). DNA libraries were generated by the Skoltech Genomics Core Facility using Swift Accel-NGS 1S Kit (Swift Biosciences) following the manufacturer's instructions.
Runs: 1 run, 2.1M spots, 313.4M bases, 125.3Mb
Run# of Spots# of BasesSizePublished
SRR253029282,061,976313.4M125.3Mb2023-12-12

ID:
28481108

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