Sequencing

11th Annual Sequencing, Finishing, and Analysis in the Future Meeting
TARGETED CRISPR/CAS9 DNA FRAGMENTATION AND
SELECTIVE PRIMER SEQUENCING ENABLES
MASSIVELY PARALLEL MICROSATELLITE ANALYSIS
Thursday, 2nd June 13:20 La Fonda Ballroom Talk (OS‐6.01)
Gi Won Shin, Susan M Grimes, Hojoon Lee, Billy Lau, Charlie Xia, Hanlee P Ji
Stanford University
Microsatellites are multi‐allelic genetic markers, composed of short tandem repeats (STRs). They
have unit motifs composed of mononucleotides, dinucleotides and large motifs up to hexamers. Next
generation sequencing approaches and other methods for STR analysis rely on the analysis of a
limited number of PCR amplicons, typically in the tens. To massively increase throughput and improve
genotyping accuracy of microsatellites, we developed STR‐Seq, a next generation sequencing
technology that analyzes over 1,000 STRs in parallel. STR‐Seq uses in vitro CRISPR/Cas9
fragmentation to produce specific DNA molecules that encompass the complete microsatellite
sequence. Afterwards, STR‐selective primers enable massively parallel, targeted sequencing of large
STR sets. Amplification‐free library preparation provides single molecule sequences without using
unique molecular barcodes. Overall, STR‐Seq’s genotyping has high throughput, high accuracy and
provides informative haplotypes. Using these new features, STR‐Seq can identify a 0.1% minor
genome fraction in a DNA mixture composed of different, unrelated samples.
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