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Sequencing dMDA Products on the MinION using Oxford Nanopore’s Rapid Barcoding Kit
Output Details
Description
Using Oxford Nanopore MinION and the Rapid Barcoding library preparation kit, we perform long read sequencing on droplet multiple displacement amplification (dMDA) products generated from single nuclei. Single nuclei contain around 6.6pg of DNA, to sequence this miniscule amount of DNA using long reads a whole genome amplification (WGA) method must be used to provide enough starting material for library preparation. MDA is currently the only WGA method which produces reads >10kb in length. Sequencing DNA of this length using long reads can allow for direct resolution and identification of most structural variants, including large LINE-1 insertions and deletions. dMDA is an improvement of the MDA technique, partitioning DNA template molecules into droplets prevents the generation of intermolecular chimeras, and a finite amount of dMDA reagents being packed into each droplet, prevents overamplification of certain reads. The expected result of this protocol is to be able to produce and sequence libraries generated from dMDA products.
Identifier (DOI)
10.17504/protocols.io.kxygxy9odl8j/v1