cuttag

1、官方流程,太空了,还是得结合文章分析

CUT&Tag Data Processing and Analysis Tutorial

2、paper流程参考

【Metabolic reprogramming by histone deacetylase inhibition preferentially targets NRF2-activated tumors】

【TASOR is a pseudo-PARP that directs HUSH complex assembly and epigenetic transposon control】

【Genome-wide chromatin accessibility is restricted by ANP32E】

【The BAF chromatin remodeler synergizes with RNA polymerase II and transcription factors to evict nucleosomes】

【CSTF2 mediated mRNA N6-methyladenosine modification drives pancreatic ductal adenocarcinoma m6A subtypes】

【Regulation of CTCF loop formation during pancreatic cell differentiation】

paperfastqcremove adapteralignment hg38/mm10

alignment

E. coli 

remove duplicates

Genomic enrichmentsPeaks
Metabolic reprogramming by histone deacetylase inhibition preferentially targets NRF2-activated tumors

Bowtie2

parameters: –local –very-sensitive-local –no-unal –no-mixed –no-discordant –phred33 -I 10 -X 700

(parameter: REMOVE_DUPLICATES = true) of Picard deeptools (v3.3.2, parameters bamCoverage –normalizeUsing CPM –binSize 25 –smoothLength 100) and visualized using IGVMACS2 (parameters: –f BAMPE -g mm –broad)
TASOR is a pseudo-PARP that directs HUSH complex assembly and epigenetic transposon control Bowtie2 (--local –very-sensitive-local –no-mixed –no-discordant -I 10 -X 700)PCR duplicates were removed using Picardtracks were generated using bamCoverageResulting bigwig files were displayed in IGV and heatmaps made with deepTools computeMatrix and plotHeatmap packages
Genome-wide chromatin accessibility is restricted by ANP32EBowtie2PCR duplicates were removed using Picardgenome browser tracks in bigwig format were generated from merged replicates using deeptools bamCoverageMACS2 (setting for H2A.Z ChIP: -f BAMPE–SPMR–nomodel -B–broad; for ATAC-seq: -f BAMPE–SPMR–nomodel -B)
The BAF chromatin remodeler synergizes with RNA polymerase II and transcription factors to evict nucleosomesadapters were clippedBowtie2  with parameters: --very-sensitive-local --soft-clipped-unmapped-tlen --dovetail --no-mixed --no-discordant -q --phred33 -I 10 -X 1000mapped to Ensembl masked R64-1-1 with parameters: --end-to-end --very-sensitive --no-overlap --no-dovetail --no-unal --no-mixed --no-discordant -q --phred33 -I 10 -X 700. Continuous-valued data tracks (bedGraph and bigWig) were generated using genomecov in bedtools v.2.30.0peaks were called by SEACR (v.1.3) using the norm and relaxed settings
CSTF2 mediated mRNA N6-methyladenosine modification drives pancreatic ductal adenocarcinoma m6A subtypes FastQCAdaptor and low-quality bases were removed using FastpBowtie2 with options: -p 6 --local --very-sensitive-local --no-unal --no-mixed --no-discordant --phred33 -I 10 -X 700Visualization of the depositions along genomic regions was performed with IGVmacs2 callpeak -t input_file -p 1e-5 -f BAMPE –n out_name
Regulation of CTCF loop formation during pancreatic cell differentiationBowtie2 and the hg38 human genome releasePCR duplicates were removed using Picard ToolsNormalized reads were used to derive bedgraph files for comparison in IGV.MACS2 was used to call peaks using default parameters with IgG ChIP-seq data as control

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