NadPrep cfDNA Library Preparation Kit (for Illumina®️)

Catalog Number:

#1002101 #1002103
#1003211 #1003212
#1003213 #1003221 #1003224
#1003240 #1003241 #1003242
#1003251 #1003252 #1003253 #1003254

NadPrep cfDNA Library Preparation Kit (forIllumina®️) is designed for preparation of high-quality libraries from double-stranded DNA (dSdna) on Illumina platforms. This A-T ligation-based kit offers a stable and efficient library preparation solution forgenome sequencing with cfDNA, andcompatible withhybridization capture based targeted sequencing onIllumina platforms.

This kit includes Unique Molecular Identifier (UMI) adapters, which can enhance the performance of mutant detection with ultralow-frequency in cfDNA.

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Features:

Applicable to cfDNA ranging from 1 ng to 500 ng
Generate high quality libraries from low input
Unique Molecular Identifier (UMI) adapter module enables accurate calling of low-frequency variants
Provided with DNA purification magnetic beads and optimized size-selection solution

Workflow

workflow

Performance

1% (0.5%) variants were mimicked by spiking plasma cfDNA from one healthy donor into that of the unrelated other at 1% proportion. 10 ng cfDNA mixture were used for UMI library preparation. Targeted SNPs were enriched by in-house panels and sequenced on Illumina Hiseq X (PE150) (Figure 1). UMIs allow the error correcting algorithms to filter out false positive calls.

Figure1. Low-frequency variant model.

Table 1. Different consensus reads filtering setting and requirements.

Methods	Description
No UMI	Mark reads with the same start-stop positions as duplicates.
SSCS	makesuse of start-stop position coupled with single strand molecule identifiers (Call Molecular Consensus Reads) to generate Single Strand Consensus Sequence (SSCS)
DCS211/ DCS633	Makes use of start-stop position coupled with double strand (top and bottom strands) molecule identifiers to generate Duplex Consensus Sequence (DCS). the top and bottom reads≥ 1(DCS211)/3(DCS633)
DCS211(≥2)/DCS633(≥2)	Based on the data of DCS211/ DCS633, the DCS of the same Variants should be ≥ 2

Consistent with the expected mutation frequency

Figure 2. The frequency estimated for 1% and 0.5% allele variation.

Denoising effect achieved by different stringency of consensus filtering.

Figure 3. Denoising effect achieved by different stringency of consensus filtering.

Sensitivity and positive predictive value

Figure4. A. Sensitivity and B. positive predictive value (PPV) for 1% and 0.5% variant.

For research use only. Not for use in diagnostic procedures.

NadPrep DNA Library Preparation Module (for Illumina)

Package	Color of Tube Cap	Component	Volume 1002101 (24 rxn)	Volume 1002103 (96 rxn)
Box1		EndRepair & A-Tailing Buffer	180 µL	690 µL
		End Repair & A-Tailing Enzyme	120 µL	460 µL
		Ligation Buffer	750 µL	2×1500 µL
		DNA Ligase	60 µL	230 µL
		2X HiFi PCR Master Mix	720 µL	2×1600 µL
		NadPrep Amplification Primer Mix	145 µL	650 µL
	/	Nuclease Free Water	2.5 mL	8 mL
		TE Solution	1 mL	4 mL
Box2	/	NadPrep SP Beads	4 mL	15 mL

Unique Molecular Identifier (UMI) adapters module,

Color of Tube Cap	Component	Volume 1103111 (24 rxn)	Volume 1103121 (96 rxn)	Volume 1103122 (96 rxn)
	NadPrep Universal UDI-Index Primer Mix #	12×15 µL	24×25 µL	24×25 µL
	NadPrep UMI Adapter	60 µL	230 µL	230 µL
	NadPrep Amplification Primer Mix II	20 μL	75 μL	75 μL

Product	Scale	Catalog
NadPrep DNA Library Preparation Kit (for Illumina^®️) G24	24 rxn	1002101
NadPrep DNA Library Preparation Kit (for Illumina^®️) E96	96 rxn	1002103
NadPrep UMI Adapter Kit Set A1 (with 10 nt Index), 24 rxn	1-12	1103111
NadPrep UMI Adapter Kit Set B1 (with 10 nt Index), 96 rxn	1-24	1103121
NadPrep UMI Adapter Kit Set B2 (with 10 nt Index), 96 rxn	25-48	1103122