ONT-LibQC
Example report
ONT LIBRARY QUALITY CONTROL

HL240

Shallow sequencing · whole-molecule library assessment

chromatinhg382,936 raw moleculesont_native · MAPQ ≥20 · 10 kb bins

Total library

ONT Sequencing QCpass
NGS Library Type / Shape QCpass
01 · SEQUENCING

ONT Sequencing QC

MetricReference intervalTotal library
Raw reads ≥500
2,936
ONT sequencing QC Local yield screen
pass
Mean read Q Chemistry-specific
19.46
Raw median length Library-dependent
276 bp
Raw read N50 Library-dependent
294 bp
Inner-anchor reconstruction rate 50.0%–100.0%
87.8%
02 · NGS LIBRARY QC

NGS Library Type / Shape QC

Library identity

Reference assemblyhg38
Assaychromatin
Library modeSingle Library
Top-pair support98.1%
A dominant i5–i7 pair is single_library at ≥90% informative support, or when it is the only supported pair. Pair support is conditional on index-informative units; inspect overall assignment and ambiguous/unassigned fractions below.

Resolved physical index pair(s)

Libraryi5 (P5 side)i7 (P7 side)Support
lib01AAGACACCGTAACCGA98.1%
P5/P7-compatible adapter family · truseq

Physical i5 on the P5 side and i7 on the P7 side; sequences are normalized to P5→P7 orientation.

MetricReference intervalTotal library
Adapter-dimer QC ≥20 units; see burden
pass
Adapter-dimer burden 0.0%–5.0%
0.1%
Insert median length Target / protocol-specific
146 bp
Both outer handles detected Both P5 and P7 expected
82.3%
All-unit index assignment Prefer high assignment
98.2%
Primary-alignment rate 50.0%–100.0%
98.3%
Mappable units Depth-dependent
2,586
Additional structure, index and reconstruction metrics
MetricReference intervalTotal library
Units with both outer handles Depth-dependent
2,131
Ambiguous index fraction Prefer 0%
0.0%
Unassigned index fraction Prefer 0%
1.8%
Unresolved dimer candidates Prefer 0; assess fraction
8
Reconstructed-unit dimer rate Prefer 0%
0.1%
Concatemer rate Preparation-dependent
0.3%
Library units Depth-dependent
2,589
Units per complete read ≥1 by definition
1.004
Canonical adapter dimers Prefer 0; assess fraction
3
Structure-matched adapter dimers Prefer 0; assess fraction
0
Raw structure-dimer rate Prefer 0%
0.1%
Reference intervals: basis and sources

Hover a reference value for its basis. Numeric colored ranges are local screening settings unless stated otherwise. Biological expectations, statistical baselines and design objectives provide context and do not introduce new pass/fail thresholds.

Adapter dimers. Illumina recommends ≤0.5% for patterned flow cells and ≤5% for non-patterned flow cells, assessed relative to the full library by trace/electropherogram analysis. These percentages are not calibrated equivalents of ONT raw-molecule dimer burden. Illumina: adapter dimers.

Library handles. Both P5 and P7 are required for cluster formation; observed ONT handle recovery also depends on sequencing and reconstruction. Illumina: library quantification.

Fragment sizes. CUT&RUN has target-dependent short and nucleosomal components. Micro-C MNase titration evaluates approximately 150-bp nucleosomal DNA and longer products before ligation. Neither provides a universal acceptable insert median or a calibrated threshold for the inferred fragments in this report. Skene & Henikoff, 2017; Mapping Mammalian 3D Genome Interactions with Micro-C-XL.

Size and adapter interpretation

The 120–170 bp raw-size window is supportive only. Dimer-like units have a ≤10-bp central gap. Unreconstructed template matches remain candidates; resolved 11–50 bp inserts are retained. Full-library sizes include expected outer adapters and inferred physical indexes, including units with undetected outer handles. The main profile weights each recovered unit by its modeled full-library length; count-based profiles are expandable. Curves are conditional on sequencing and adapter recovery and do not measure TapeStation fluorescence or NGS molarity.

Library profiles · size & genomic distribution

Total library

Full library = insert + 136 bp adapter/index sequence. 2,589 modeled units. The main curve weights each unit by its full-library length.

Download both profiles · Modeled unit lengths · Model details

Count-based profiles · full library, raw reads and inserts

Each recovered unit contributes one count to the full-library and insert profiles; each raw read contributes one count to the raw profile. Each curve has its own peak set to 100.

Chromosome coverage · auxiliary diagnostic

Inspect broad coverage imbalance and isolated concentration. Gene density and shallow coverage alone cannot establish target-specific enrichment or copy-number changes.

Mapped reads by chromosome karyotype
03 · ASSAY EVIDENCE

Chromatin-specific QC

MetricReference intervalTotal library
Collision O/E (depth normalized) 1 = uniform-null baseline
6.798
Contributing ONT molecules Evidence count
2,185
Usable fragments Depth-dependent
2,195
Usable fragment rate 30.0%–100.0%
84.9%
Mitochondrial fraction 0.0%–20.0%
0.1%
Mycoplasma signal screen NOT_DETECTED
NOT_DETECTED
Mycoplasma-screened inserts Depth-dependent
2,586
Mycoplasma-supporting reads 0
0
Mycoplasma-supporting loci 0
0
Mycoplasma signal fraction 0%
0.0%
Assay interpretation

Chromatin enrichment is not classified. Collision O/E is depth normalized (random baseline 1). Sampling uncertainty remains at low depth. TSS–TES profiles are CPM normalized and visual only.

TARGET CONTEXT

Custom region metaplot

Choose a BED file for hg38. The profile is calculated locally in this browser.

Normalization and region handling

BED uses zero-based, half-open coordinates. BED6 minus strands are reversed; BED3/4 and dot strands use genomic orientation. Auto uses reference point mode when all retained intervals are 1 bp; otherwise it uses region mode, including mixed-width BED files. You can override this with Plot mode. Reference point mode centers each interval at its midpoint (start + end)/2 and shows ±5 kb in 100 bins without scaling a body. Region mode scales bodies to 100 bins with 50 bins across each 5 kb flank. Exact aligned-base coverage (CIGAR M/= /X only; D/N excluded) is divided by each bin's available genomic width, then CPM uses all eligible primary nuclear alignments in the selected scope. Regions are equally weighted. Duplicate intervals are removed; non-primary and absent-contig regions are skipped and counted. Flanks are clipped to chromosome boundaries; bins entirely beyond the chromosome are missing. SEM is across regions. Assembly identity must be confirmed; chromosome names alone cannot verify it. No data are uploaded to a server.

REFERENCE SCREENING

Automatic spike-in candidate screen ?

2586 screened inserts · No spike-in declared

Reference candidateRoleInsertsFractionSupported lociResult
Saccharomyces cerevisiaespikein candidate00.0%0
NOT_DETECTED
Schizosaccharomyces pombespikein candidate00.0%0
NOT_DETECTED
Lambda phagesequencing control00.0%0
NOT_DETECTED
PhiX174sequencing control00.0%0
NOT_DETECTED
Escherichia colispikein candidate00.0%0
NOT_DETECTED
Drosophila melanogasterspikein candidate00.0%0
NOT_DETECTED

Per-unit assignments · Screen details