NadPrep ES Hybrid Capture Reagents

Catalog Number:

#1005401
#1005402

NadPrep ES Hybrid Capture Reagents is a kit optimized for the targeted capture with rapid hybridization elution steps of NAD Panels/Probes developed by Nanodigmbio. The complete liquid-phase hybridization capture system is composed of NadPrep NanoBlockers series and 120 nt NAD Panels/Probes series, and supports flexible selection between 0.5-4 hr rapid hybridization and overnight hybridization by adjusting the hybridization time to optimize the capture performance for panels of different sizes.

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Feature

● Simplify the types of elution buffer into one to reduce the possibility of human errors during operation

● Simplify the experimental process, make it more convenient and user-friendly for operation to facilitate the development of automatic workstation.

●Support flexible selection between rapid hybridization for 0.5-4 hr or overnight hybridization for 16 hr.

● Support the adjustment of hybridization time to optimize capture performance.

Performance

Capture performance with various hybridization time

Fig 1.Capture performance of NadPrep ES Hybrid Capture Reagents with various hybridization time.100 ng of Human Genomic DNA (Promega, G1471) was used for pre-library preparation with the NadPrep EZ DNA Library Preparation Kit v2, with 25 min of enzymatic digestion at 25°C; 500 ng/pre-library (1-plex) input was used, following the user manual of hybrid capture of NadPrep ES Hybrid Capture Reagents (0.5/1/2/4/16 hr_ES) and NadPrep Hybrid Capture Reagents (16 hr_Trad). Sequencing was performed using Illumina Novaseq 6000, PE 150 and DNBSEQ-T7, PE 150. Appropriate amount of data was selected for subsequent analysis. The BWA was used for alignment to the reference genome hg38 and On-target rate was calculated by the number of reads.。 A. On-target rate；B. Fold 80 base penalty；C. 0.2x mean depth；D. 0.5x mean depth。
Note：41.7 Mb, 2.6 Mb and 0.23 Mb in the assay are NEXome Core Panel, NanOnco Plus Panel v2.0 and LungCancer Panel v1.0, respectively.

Capture performance for panels of different sizes

Fig 2. Capture performance of NadPrep ES Hybrid Capture Reagents with 2 hr for hybridization using panel of different size.100 ng of Human Genomic DNA (Promega, G1471) was used for pre-library preparation with the NadPrep EZ DNA Library Preparation Kit v2, with 25 min of enzymatic digestion at 25°C; 500 ng/pre-library (1-plex) input was used, following the user manual of hybrid capture of NadPrep ES Hybrid Capture Reagents. Sequencing was performed using DNBSEQ-T7, PE 150. Appropriate amount of data was selected for subsequent analysis. The BWA was used for alignment to the reference genome hg38 and On-target rate was calculated by the number of readsA. On- target rate and Mappability；B. Target covered；C. Fold 80 base penalty and Duplication。

Note： 3.27 Mb, 0.38 Mb and 0.028 Mb in the assay are HiSNP Plus Panel v1.0, NanOnCT Panel v1.0 and NanoID Panel v1.0, respectively, and the rest param- eters are the same as above.

Capture performance on different sequencing platforms

Figure 3. Capture performance of NadPrep ES Hybrid Capture Reagents on different platforms. 1100 ng of Human Genomic DNA (Promega, G1471) was used for pre-library preparation with the NadPrep EZ DNA Library Preparation Kit v2, with 25 min of enzymatic digestion at 25°C; 500 ng/pre-library (1-plex) input was used, following the user manual of hybrid capture of NadPrep ES Hybrid Capture Reagents. Sequencing was performed using Illumina Novaseq 6000, PE 150 and DNBSEQ- T7, PE 150. Appropriate amount of data was selected for subsequent analysis. The BWA was used for alignment to the reference genome hg38 and On-target rate was calculated by the number of reads. A. On-target rate and Mappability;；B. Target covered；C. Fold 80 base penalty and Duplication；D. The unbiased of GC coverage.
Note：The unbiased of GC coverage is only presented as 2.6 Mb Panel.

Multiple variants analysis SNV EGFR L858R SNV KRASA146T SNV NRAS Q61K SNV EGFR T790M

Fig 4. Correlation between expected allele frequency and observed allele frequency. 50 ng of PancancerLight 800 gDNA Reference Standard (Genewell, GW-OGTM800) was used for pre-library preparation with the NadPrep EZ DNA Library Preparation Kit v2, with 25 min of enzymatic digestion at 25°C; 500 ng/pre- library (1-plex) input was used, following the user manual of hybrid capture of NadPrep ES Hybrid Capture Reagents and NadPrep Hybrid Capture Reagents, using the 0.23 Mb panel for 1 hr rapid hybridization (1 hr_ES) and 16 hr overnight hybridization (16 hr_Trad.). Sequencing was performed using Illumina Novaseq 6000, PE 150 and DNBSEQ-T7, PE 150. Appropriate amount of data was selected for variant analysis using Vardict.

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

Package	Color Of Tube Cap	Component	Volume 1005402 (16 rxn)	Volume 1005401 (96 rxn)
Package	Color Of Tube Cap	Component	Volume 1005402 (16 rxn)	Volume 1005401 (96 rxn)
Box 1		ES Hyb #1	380 μL	2×1150 μL
		Hyb #2	120 μL	720 μL
		Human Cot DNA	110 μL	650 μL
	/	ES Wash Buffer	4×4 mL	5×19 mL
		Sealing Oil	250 μL	1.5 mL
Box 2	/	Streptavidin Beads	950 μL	5.7 mL