scRC-Seq

Single-Cell Retrotransposon Capture Sequencing

Single-cell RC-seq uses sequence capture to enrich DNA for the junctions between retrotransposon termini and adjacent genomic regions, followed by paired-end sequencing (Upton et al., 2015). In this method, nuclei are purified first by fluorescence-activated cell sorting (FACS) and then picked with a micromanipulator. The DNA is extracted from nuclei and subjected to linear WGA, followed by exponential PCR in 2 separate reactions for each nucleus, using different enzymes. The products are combined and analyzed via WGS to assess genome coverage. The libraries prepared are enriched by hybridization to locked nucleic acid (LNA) probes and sequenced (Sun et al., 2015)

Advantages:

• Detects somatic mutations in single cells

Disadvantages:

• Low throughput due to manual single-nucleus isolation

Reagents:

Illumina Library prep and Array Kit Selector



Reviews:

None available yet

References:

Upton K. R., Gerhardt D. J., Jesuadian J. S., Richardson S. R., Sanchez-Luque F. J., et al. Ubiquitous L1 mosaicism in hippocampal neurons. Cell. 2015;161:228-239

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History: scRC-Seq

Revision by sbrumpton on 2017-06-21 07:50:21 - Show/Hide