chip seq library preparation (Novogene)
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Novogene
chip seq library preparation
Chip Seq Library Preparation, supplied by Novogene, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/chip+seq+library+preparation/chip+library+preparation+seq/bio_rxiv__64898__2026__02__23__707304-115-5-9
Average 86 stars, based on 1 article reviews
Chip Seq Library Preparation, supplied by Novogene, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/chip+seq+library+preparation/chip+library+preparation+seq/bio_rxiv__64898__2026__02__23__707304-115-5-9
Average 86 stars, based on 1 article reviews
chip seq library preparation - by Bioz Stars,
2026-09
86/100 stars
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1) Product Images from "Modulation of rob expression accelerates development of antibiotic resistance in Yersinia enterocolitica"
Article Title: Modulation of rob expression accelerates development of antibiotic resistance in Yersinia enterocolitica
Journal: bioRxiv
doi: 10.64898/2026.02.23.707304
Figure Legend Snippet: (A–D) ChIP-seq profiles showing Rob binding at the promoter regions of meoA (A), tolC (B), mlaF (C), and atpI (D). Blue tracks represent normalized ChIP-seq signal in the control strain (top) and Rob-tagged strain (bottom). Gene orientations are indicated by arrows; scale bar, 500 bp. (E) Schematic model of Rob-dependent transcriptional regulation. Rob binding upstream of target operons activates genes involved in outer membrane permeability ( ompF/ompD ), multidrug efflux ( tolC–acrAB2 ), and phospholipid transport ( mla operon). (F) Relative expression of selected Rob regulon genes measured by qPCR, shown as fold change normalized to wild type (WT). Data represent mean ± SD. The red dashed line indicates WT expression level. (G) Corresponding fold changes in gene expression derived from RNA-seq analysis, normalized to WT, confirming global upregulation of Rob target genes. (H) Efflux activity measured over time using a fluorescence-based assay. Rob mutant exhibits significantly higher efflux compared to the WT. Data are shown as mean ± SD; **** indicates P < 0.0001.
Techniques Used: ChIP-sequencing, Binding Assay, Control, Membrane, Permeability, Expressing, Gene Expression, Derivative Assay, RNA Sequencing, Activity Assay, Fluorescence, Mutagenesis
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Immunoprecipitation:Article Title: Modulation of rob expression accelerates development of antibiotic resistance in Yersinia enterocolitica Article Snippet: Followed by transcription initiation blocking with rifampicin (final 150 mg/L; stock 50 mg/mL in methanol, 300 μL per sample) for 20 min at 28 °C, 220 rpm; cells were crosslinked by adding 37% formaldehyde to 1% final for 20 min at 28 °C, 220 rpm, and the reaction was quenched by adding 2.5 M glycine to 125 mM final, incubating 20 min at 4 °C with gentle shaking; cells were pelleted (6000×g, 5 min, 25 °C), washed three times with 1× PBS, and pellets were processed immediately or stored at −80 °C; pellets were resuspended in 200 μL lysozyme (20 mg/mL) and incubated 30 min at 37 °C, chromatin was fragmented by sonication (Covaris, M220 Focused-ultra-sonicator) for 3 minutes to generate DNA fragments of approximately 150–400 bp, debris were removed (14,000 rpm, 10 min); lysates were pre-cleared with 50 μL Protein A/G beads (MedChemExpress) for 2 h at 4 °C on a rotating wheel, beads were pelleted (13,800×g, 1 min) and the supernatant was incubated with 10 μL mouse anti-FLAG antibody overnight at 4 °C with rotation, then immune complexes were captured with 50 μL Protein A/G beads for 6 h at 4 °C (or overnight) and beads were collected (10,000 rpm, 5 min), washed four times with lysis buffer (NaCl, HEPES, sodium deoxycholate, Triton X-100, DTT, protease inhibitor cocktail 1:100, and Tris-HCl) at 4 °C (1 mL each; 14,000 rpm, 5 min) and twice with TE buffer; complexes were eluted with 100 μL elution buffer 1 at 65 °C for 10 min and then 150 μL elution buffer 2, pooling eluates (250 μL total), and crosslinks were reversed at 65 °C for 6 h or overnight; 10% eluate (25 μL) was reserved for optional Western blot, the remainder was treated with RNase A (20 μL; 100 mg/L) for 30 min at 37 °C and digested with proteinase K (250 μL of a proteinase K/glycogen mix prepared in TE) for 2 h at 58 °C, followed by DNA cleanup with 4 M LiCl (55 μL), phenol:chloroform:isoamyl alcohol extraction (25:24:1) and ethanol precipitation (−80 °C, 30 min; 14,000 rpm, 5 min, 4 °C), washing with 70% ethanol, air-drying, and resuspension in 25 μL nuclease-free water; DNA was quantified (Qubit) and stored at −20 °C. .. Immunoprecipitated DNA was subjected to Chromatin Immunoprecipitation:Article Title: Modulation of rob expression accelerates development of antibiotic resistance in Yersinia enterocolitica Article Snippet: Followed by transcription initiation blocking with rifampicin (final 150 mg/L; stock 50 mg/mL in methanol, 300 μL per sample) for 20 min at 28 °C, 220 rpm; cells were crosslinked by adding 37% formaldehyde to 1% final for 20 min at 28 °C, 220 rpm, and the reaction was quenched by adding 2.5 M glycine to 125 mM final, incubating 20 min at 4 °C with gentle shaking; cells were pelleted (6000×g, 5 min, 25 °C), washed three times with 1× PBS, and pellets were processed immediately or stored at −80 °C; pellets were resuspended in 200 μL lysozyme (20 mg/mL) and incubated 30 min at 37 °C, chromatin was fragmented by sonication (Covaris, M220 Focused-ultra-sonicator) for 3 minutes to generate DNA fragments of approximately 150–400 bp, debris were removed (14,000 rpm, 10 min); lysates were pre-cleared with 50 μL Protein A/G beads (MedChemExpress) for 2 h at 4 °C on a rotating wheel, beads were pelleted (13,800×g, 1 min) and the supernatant was incubated with 10 μL mouse anti-FLAG antibody overnight at 4 °C with rotation, then immune complexes were captured with 50 μL Protein A/G beads for 6 h at 4 °C (or overnight) and beads were collected (10,000 rpm, 5 min), washed four times with lysis buffer (NaCl, HEPES, sodium deoxycholate, Triton X-100, DTT, protease inhibitor cocktail 1:100, and Tris-HCl) at 4 °C (1 mL each; 14,000 rpm, 5 min) and twice with TE buffer; complexes were eluted with 100 μL elution buffer 1 at 65 °C for 10 min and then 150 μL elution buffer 2, pooling eluates (250 μL total), and crosslinks were reversed at 65 °C for 6 h or overnight; 10% eluate (25 μL) was reserved for optional Western blot, the remainder was treated with RNase A (20 μL; 100 mg/L) for 30 min at 37 °C and digested with proteinase K (250 μL of a proteinase K/glycogen mix prepared in TE) for 2 h at 58 °C, followed by DNA cleanup with 4 M LiCl (55 μL), phenol:chloroform:isoamyl alcohol extraction (25:24:1) and ethanol precipitation (−80 °C, 30 min; 14,000 rpm, 5 min, 4 °C), washing with 70% ethanol, air-drying, and resuspension in 25 μL nuclease-free water; DNA was quantified (Qubit) and stored at −20 °C. .. 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Immunoprecipitated DNA was subjected to Purification:Article Title: Multi-omics dissection of the genetic regulation underlying fatty acid composition in cattle Article Snippet: DNA concentration was measured using Qubit® DNA Assay Kit in Qubit® 3.0 Fluorometer (Life Technologies, CA, USA). .. The purified DNA was used for Construct:Article Title: Multi-omics dissection of the genetic regulation underlying fatty acid composition in cattle Article Snippet: DNA concentration was measured using Qubit® DNA Assay Kit in Qubit® 3.0 Fluorometer (Life Technologies, CA, USA). .. The purified DNA was used for |