Rapid Hyb | NadPrep ES Hybrid Capture Reagents Background Liquid-phase hybridization capture sequencing is, based on the principle of complementary base pairing, to design and synthesize nucleic acid probes for regions of interest, so that DNA libraries can be enriched and sequenced using liquid-phase hybridization in target regions, so as to achieve detection on target regions at lower costs. Thanks to the ability of the probes for tolerating certain differences between target regions and the ability for jointly capturing surrounding regions, liquid-phase hybridization capture can accurately detect a variety of mutations such as SNV, InDel, CNV, SV, and gene fusions. Due to its wide application in supporting disease exploration, the detection of new pathogens or pathogenic genes has been greatly improved. As a cost-effective, reproduci- ble and stable research strategy, it is also of great significance for personalized treatment of specific individual genetic backgrounds. More rapid and stable experimental workflows are urgently needed for the detection of both tumor and pathogenic microorganism infections, so as to further streamline the process. Overnight hybridization (~16 hr) and redundant elution in traditional liquid-phase hybridization capture have undoubtedly affect the detection process. Therefore, Nanodigmbio has launched NadPrep ES Hybrid Capture Reagents (ES, Excellent Speed), which can be compatible with rapid hybridization capture of different probes, so as to accelerate the detection process. 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. 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. Vacuum concentration Vacuum concentration Bead Wash Buffer Vacuumconcentratio Hybrid Beads wash Capture Wash PCR Purification Note: Nad-Traditional indicates NadPrep Hybrid Capture Reagents; NadPrep ES indica

Capture performance with various hybridization time A 100 Fold 80 base penalty Figure 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℃; 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...

Capture performance for panels of different sizes A Figure 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℃; 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...

Capture performance on different sequencing platforms A Target covered (%) Normalized coverage Fold 80 base penalty Figure 3. Capture performance of NadPrep ES Hybrid Capture Reagents on different platforms. 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℃; 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...

Multiple variants analysis Observed allele frequency (%)/copy number Type Variant Reference 16 hr_Trad. 1 hr_ES Insertion EGFR V769_D770 insASV 3% SNV EGFR G719S 4% Observed allele frequency Figure 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/prelibrary (1-plex) input was used, following the user manual of hybrid capture of NadPrep ES Hybrid Capture...