在准备NGS文库的时候,会有用到转座酶Tn5,Nextera DNA Library Preparation Kit,比如ATAC-seq就有用到这个Tn5。转座酶携带有特定的序列称为转座子Transposon。 下面是ATAC-seq的工作原理,想必听说过ATAC-seq的,对这个图都会再熟悉不过了。 Tn5是kit里面带有的,没什么大不同。ATAC-seq 的barcoded primers (adaptors...
Illumina Nextera DNA Library Preparation Kit 研究人员发现整个转座子序列并不是转座必须的,只需转座子的末端核心序列,转座酶便能将该部分序列插入并连接至基因组内;根据这个原理,将测序接头序列加入末端核心序列中,可简捷地引入测序接头,完成文库构建。 由下图可看到在tn5二聚体里面的双链Oligo是Nextera Tn5 bindi...
Tn5 enzyme in this kit is a hyperactive mutant that we cloned, expressed, purified, and test in-house. The Assembled Transposomes containing this enzyme are provided pre-loaded with the next-gen sequencing adapters, so it is ready-to-use without any optimization or other preparation required. ...
The optimized protocol guides you through the steps for sample preparation, tagmentation and library preparation, yielding next-gen sequencing-ready libraries that can be multiplexed in a single flow cell sequencing run. ATAC-Seq Kit Highlights: Reveal the genomic sequence of open chromatin regions ...
RNA isolation, RNA-Seq library preparation, and sequencing RNA was extracted according to the manufacturer’s instructions using the RNeasy Kit (QIAGEN). For RNA-seq Illumina TruSeq Stranded Total RNA Library Preparation Kit was used using 1000 ng RNA as input. Sequencing libraries were quantifi...
Nextera DNA Library Preparation Kit (2x TD Buffer, Tn5 Enzyme) Illumina FC1211030 Critical Component Nuclease Free Sterile dH20 Gibco 10977015 Can use other comparable products. Polysorbate 20 (Tween20) Sigma P9416 Can use other comparable products. PowerUp SYBR Green Master Mix Applied Biosystems...
(Phased-txci-ATAC-seq) that effectively decouples sample processing from library preparation. Specifically, on the sample processing day, nuclei are isolated, quantified, and then cryopreserved in either PCR tube strips or a 96-well plate. On the designated library preparation day, the frozen ...
Library preparation used the TrueSeq Stranded Total RNAseq Poly A1 Gold Kit (Illumina), according to the manufacturer’s recommendations, followed by mRNA enrichment using poly(A) enrichment for ribosomal RNA removal. Sequencing was performed using the HiSeq platform (Illumina) for 2 × 100...
ATAC-seq (Assay for Transposase-Accessible Chromatin with high-throughput sequencing) provides an efficient way to analyze nucleosome-free regions and has been applied widely to identify transcription factor footprints. Both applications rely on the accu
Investigators were not blinded to allocation during library preparation, experiments, or analysis. Reporting summary Further information on research design is available in the Nature Portfolio Reporting Summary linked to this article.Data availability All datasets used in this work are publicly available. ...