A ligation-based combinatorial barcoding strategy introduces second and third rounds of DNA barcodes nuclei are pooled and separated into wells creating unique barcodes for each nucleus.Tagmentation (cleavage and barcode tagging of chromatin DNA and cDNA by Tn5) and reverse transcription are performed.Antibodies against a specific histone modification are used to target Tn5 transposase to chromatin.The Paired-Tag protocol can be broken down into 4 key steps:.
By adapting the tagmentation and cleavage steps, they have optimized the approach to pair single-cell ChIP-seq and RNA-seq. This method works by ligating specific barcodes to open chromatin and cDNA from each cell, followed by sequencing. In their latest, the group focused on extending a technique they previously developed ( Paired-seq), which pairs chromatin accessibility and RNA-seq in single cells. Previously, the Ren lab has pioneered chromatin conformation capture technologies, adapting some for use in single-cells. To meet this challenge, the lab of Bing Ren at the University of California San Diego developed a new technique they call Paired-Tag (parallel analysis of individual cells for RNA expression and DNA from targeted tagmentation by sequencing). While multiomic methods have been gaining momentum, it has been tricky for researchers to play matchmaker with ChIP-seq at the single-cell level. With Valentines day just behind us, a new genome-wide assay has reinforced the perks of pairing up for single-cells.
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