The Science Behind Reducing Amplification Bias
LockSeq captures and circularizes individual DNA molecules before amplification, preserving molecular identity from the start. By using a single probe per target, rather than multiple primers or overlapping probes, LockSeq simplifies target capture and reduces sources of bias and dropout
Three Steps to Accurate Target Capture
STEP 1 — HYBRIDIZATION
The padlock probe binds to flanking regions of the target site, leaving a intentional gap at the desired genomic 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 error-corrected consensus reads (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 any amplification. Reads sharing the same UMI are collapsed into a single consensus call, correcting for PCR-generated duplicates and sequencing errors (Hiatt et al. 2013).
The result: a minimized noise 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 gene editing confirmation at scale
Data Integrity
Minimizes dropouts and delivers uniform target coverage
Panel Flexibility
Modular panel design compatibility enables flexibility across development stages
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