scATAC-Seq (Cell index variation)
Single-Cell Assay for Transposase-Accessible Chromatin
This version of scATAC-seq uses combinatorial cellular indexing to measure chromatin accessibility in thousands of single cells per assay. This method avoids the need for compartmentalization of individual cells, which makes the system scalable to analyze thousands of cells at a time (Cusanovich et al., 2015).
In this method, nuclei are isolated and molecularly tagged in bulk with barcoded Tn5 transposases in wells. Next, the nuclei are pooled and a limited number redistributed into a second set of wells. A second barcode is introduced during the PCR amplification step, and the fragments are pooled for library preparation and sequencing.
Advantages:
- High throughput and scalable
Disadvantages:
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
Liu S. and Trapnell C. Single-cell transcriptome sequencing: recent advances and remaining challenges. F1000Res. 2016;5:
Lu F., Liu Y., Inoue A., Suzuki T., Zhao K. and Zhang Y. Establishing Chromatin Regulatory Landscape during Mouse Preimplantation Development. Cell. 2016;165:1375-1388
References:
Corces M. R., Buenrostro J. D., Wu B., et al. Lineage-specific and single-cell chromatin accessibility charts human hematopoiesis and leukemia evolution. Nat Genet. 2016;48:1193-1203
Cusanovich D. A., Daza R., Adey A., et al. Multiplex single cell profiling of chromatin accessibility by combinatorial cellular indexing. Science. 2015;348:910-914
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History: scATAC-Seq (Cell index variation)
Revision by sbrumpton on 2017-06-21 07:50:21 - Show/Hide
Single-Cell Assay for Transposase-Accessible Chromatin
This version of scATAC-seq uses combinatorial cellular indexing to measure chromatin accessibility in thousands of single cells per assay. This method avoids the need for compartmentalization of individual cells, which makes the system scalable to analyze thousands of cells at a time (Cusanovich et al., 2015).
In this method, nuclei are isolated and molecularly tagged in bulk with barcoded Tn5 transposases in wells. Next, the nuclei are pooled and a limited number redistributed into a second set of wells. A second barcode is introduced during the PCR amplification step, and the fragments are pooled for library preparation and sequencing.
Advantages:- High throughput and scalable
Disadvantages:Reagents:Illumina Library prep and Array Kit SelectorReviews:Liu S. and Trapnell C. Single-cell transcriptome sequencing: recent advances and remaining challenges. F1000Res. 2016;5:Lu F., Liu Y., Inoue A., Suzuki T., Zhao K. and Zhang Y. Establishing Chromatin Regulatory Landscape during Mouse Preimplantation Development. Cell. 2016;165:1375-1388References:Corces M. R., Buenrostro J. D., Wu B., et al. Lineage-specific and single-cell chromatin accessibility charts human hematopoiesis and leukemia evolution. Nat Genet. 2016;48:1193-1203Cusanovich D. A., Daza R., Adey A., et al. Multiplex single cell profiling of chromatin accessibility by combinatorial cellular indexing. Science. 2015;348:910-914