Laboratory Practices & Techniques to Maximize NGS Sample Security
Shannon Piehl
Monday, December 18, 2017
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Chapters
Introduction
Where Contamination Can Happen
MISIDENTIFICATION HAPPENS
Library Prep Can Introduce Sample Ambiguity
NEXTFLEX® RAPID DNA-SEQ KIT 2.0 Protocol
Sequencing workflow overview
Common Indexing Strategies
Edit Distance Matters
Traditional Index Design
Combinatorial Dual Indexing
Risks In Combinatorial Dual Indexing
Index Hopping On Patterned Flow Cells
Nextflex
®
Unique Dual-Indexed Barcodes
Index Misidentification On The Illumina
®
Miseq
®
Platform
Index Misidentification On The Illumina
®
Hiseq
®
4000 Platform
Unique Dual Indexes Increase Confidence In Demultiplexing
Index Misassignment In PCR-Free Applications
Unique Dual Indexes Increase Correct Index Assignment In PCR-Free Library Prep
Increased Stringency Of Demultiplexing Causes Lost Reads
Conclusions: UDIs
Conclusions
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