Standardized Tagmentase Formulation Demonstrates Consistent Performance Across Independently Manufactured Lots
To demonstrate lot-to-lot reproducibility, five independently manufactured lots of unloaded Tagmentase (Tn5 transposase) were evaluated. For each lot, transposome assembly and dilution to the required activity were performed independently according to the recommended Hologic Diagenode protocol. ATAC-seq libraries were generated from 50,000 K562 cells using the Hologic Diagenode ATAC-seq protocol and five independently manufactured Tagmentase lots, designated Lots A–E, with three replicate libraries prepared per lot. To minimize sequencing-related variability, all libraries were sequenced in a single run.
Across all five lots, ATAC-seq performance was highly consistent. Key quality metrics remained comparable, with mapping efficiencies of 98.3–98.5%, uniquely mapped reads of 76.6–77.9%, 160,000–194,000 detected peaks, FRiP values of 0.51–0.53, and average peak widths of 608–635 bp.
These results demonstrate robust and reproducible ATAC-seq library generation across independently manufactured Hologic Diagenode Tagmentase lots.
Standardized Tagmentase formulation delivers highly reproducible ATAC-seq performance
C.
Consistent ATAC-seq performance across independently manufactured standardized Tagmentase (Tn5 transposase) lots.
(A) Lot-to-lot variability of key ATAC-seq quality metrics, expressed as coefficient of variation (CV). All evaluated metrics showed CV values below 10%, indicating low manufacturing variability across Tagmentase (Tn5 transposase) production lots. (B) Pairwise Pearson correlation analysis of ATAC-seq libraries generated with independent Tagmentase (Tn5 transposase) lots. Correlation coefficients greater than 0.99 demonstrate highly similar genome-wide accessibility profiles across lots. (C) Transcription start site (TSS) enrichment profiles showing comparable chromatin accessibility patterns and signal enrichment at TSSs for all Tagmentase (Tn5 transposase) lots. (D) Representative genome browser (IGV) tracks illustrating highly similar accessibility peaks and signal intensity across independent Tagmentase (Tn5 transposase) lots. Together, these results demonstrate robust lot-to-lot reproducibility and consistent chromatin accessibility profiling using the standardized Tagmentase (Tn5 transposase) formulation.
