The Science Behind Zero Amplification Bias
Multiplex PCR amplifies everything — and loses targets in the process. LockSeq circularizes individual molecules before amplification, preserving the identity of every captured fragment. That's the difference between coverage and confidence.
Three Steps to Sequence-Level Truth
STEP 1 — HYBRIDIZATION
The padlock probe binds to flanking regions of the target site, leaving an intentional gap at the edit locus (Hardenbol et al. 2005).
STEP 2 — GAP-FILL LIGATION
DNA polymerase fills the gap. DNA ligase seals the nick — but only if base pairing is exact. The result: a circular molecule representing a single original genomic fragment, uniquely tagged with a UMI.
STEP 3 — AMPLIFICATION & SEQUENCING
Circles are amplified and sequenced. UMI groups are collapsed into consensus reads. Stochastic errors cancel out (Hiatt et al. 2013). Rare variants — down to 0.1% VAF — stand out clearly.
UMI-Native Consensus Calling
Every molecule captured by a LockSeq probe carries a unique barcode — its UMI — applied before amplification. Reads sharing the same UMI are collapsed into a single consensus call, eliminating PCR-generated duplicates and stochastic sequencing errors (Hiatt et al. 2013).
The result: a noise level low enough to resolve variants at 0.1% VAF with high confidence, across panels of 1,000+ sites.
LockSeq Enables High-Fidelity NGS-Based On- and Off-Target Confirmation
Accuracy
Enables quick and accurate on- and off-target confirmation at scale
Data Integrity
Minimizes dropouts and delivers uniform target coverage
Panel Flexibility
Supports a standardized workflow with panel flexibility across development stages
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