ssvQC: an integrated CUT&RUN quality control workflow for histone modifications and transcription factors.

ssvQC: an integrated CUT&RUN quality control workflow for histone modifications and transcription factors.
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DOI:
10.1186/s13104-021-05781-8
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发表时间:
2021-09-20
期刊:
影响因子:
1.8
通讯作者:
Frietze S
Frietze S
中科院分区:
其他
文献类型:
--
作者:
Boyd J;Rodriguez P;Schjerven H;Frietze S

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在分析细胞和组织中转录因子结合和组蛋白修饰的全基因组模式的不同方法中,CUT&RUN 已成为一种更有效的方法,与 ChIP-seq 相比,它可以使用更少数量的细胞获得更高的信噪比。 CUT&RUN 和其他相关序列富集测定的结果需要全面的质量控制 (QC) 和重复数据质量的比较分析。虽然目前存在多种用于读取映射和分析的计算工具,但缺乏数据质量的系统报告。我们的目标是 (1) 比较使用冷冻细胞和新鲜细胞进行 CUT&RUN 的方法,以及 (2) 开发一个易于使用的管道来评估数据质量。我们将 CUT&RUN 的工作流程与新鲜和冷冻样品进行了比较,并提出了一个名为 ssvQC 的 R 包,用于质量控制以及比较源自 CUT&RUN 和其他基于富集的序列数据的数据质量。使用 ssvQC,我们评估新鲜和冷冻组织样本中转录因子和组蛋白修饰的不同 CUT&RUN 方案的结果。总体而言,此过程有助于评估跨数据集的数据质量,并允许检查峰值调用分析、不同数据类型的重复分析。 ssvQC 包可在 https://github.com/FrietzeLabUVM/ssvQC 上轻松获得。在线版本包含可在 10.1186/s13104-021-05781-8 获取的补充材料。
Among the different methods to profile the genome-wide patterns of transcription factor binding and histone modifications in cells and tissues, CUT&RUN has emerged as a more efficient approach that allows for a higher signal-to-noise ratio using fewer number of cells compared to ChIP-seq. The results from CUT&RUN and other related sequence enrichment assays requires comprehensive quality control (QC) and comparative analysis of data quality across replicates. While several computational tools currently exist for read mapping and analysis, a systematic reporting of data quality is lacking. Our aims were to (1) compare methods for using frozen versus fresh cells for CUT&RUN and (2) to develop an easy-to-use pipeline for assessing data quality. We compared a workflow for CUT&RUN with fresh and frozen samples, and present an R package called ssvQC for quality control and comparison of data quality derived from CUT&RUN and other enrichment-based sequence data. Using ssvQC, we evaluate results from different CUT&RUN protocols for transcription factors and histone modifications from fresh and frozen tissue samples. Overall, this process facilitates evaluation of data quality across datasets and permits inspection of peak calling analysis, replicate analysis of different data types. The package ssvQC is readily available at https://github.com/FrietzeLabUVM/ssvQC. The online version contains supplementary material available at 10.1186/s13104-021-05781-8.
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