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ScGET-seq
Single-cell genome and epigenome by transposases sequencing (scGET-seq) is a DNA sequencing method for profiling open and closed chromatin. In contrast to single-cell ATAC-seq, assay for transposase-accessible chromatin with sequencing (scATAC-seq), which only targets active euchromatin, scGET-seq is also capable of probing inactive heterochromatin. This is achieved through the use of TnH, which is created by linking the chromodomain (CD) of Heterochromatin protein 1, heterochromatin protein-1-alpha (HP-1\alpha) to the Tn5 transposase. TnH is then able to target histone 3 lysine 9 trimethylation (H3K9me3), a marker for heterochromatin. Akin to RNA velocity, which uses the ratio of RNA splicing, spliced to unspliced RNA to infer the kinetics of changes in gene expression over the course of cellular development, the ratio of TnH to Tn5 signals obtained from scGET-seq can be used to calculate chromatin velocity, which measures the dynamics of chromatin accessibility over the course ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
ATAC-seq
ATAC-seq (Assay for Transposase-Accessible Chromatin using sequencing) is a laboratory technique used in molecular biology to assess genome-wide chromatin, chromatin accessibility. The technique was first described in 2013 as an alternative approach to MNase-seq, FAIRE-Seq and DNase-Seq but providing faster turnaround time, simplified protocol, and lower DNA input amount. Procedure ATAC-seq identifies accessible DNA regions by probing open chromatin with hyperactive mutant Transposase#Transposase Tn5, Tn5 Transposase that inserts sequencing adapters into open regions of the genome. While naturally occurring transposases have a low level of activity, ATAC-seq employs the mutated hyperactive transposase. In a process called "tagmentation", Tn5 transposase cleaves and tags double-stranded DNA with sequencing adaptors in a single enzymatic step. The tagged DNA fragments are then purified, Polymerase chain reaction, PCR-amplified, and sequenced using massive parallel sequencing, next-ge ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
