A method and a kit for generating nucleic acid for target capture

The use of circ-oligos for hybridization and RCA amplification addresses inefficiencies in existing methods, enabling efficient and cost-effective nucleic acid capture, especially in high GC-rich regions, for targeted genome sequencing.

WO2026146514A1 Publication Date: 2026-07-09TATA MEMORIAL CENTRE-ADVANCED CENTRE FOR TREATMENT RESEARCH & EDUCATION IN CANCER (TMC-ACTREC)
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Patent Information

Application Number
PCT/IN2025/051311
Authority / Receiving Office
WO · WO
Patent Type
Applications
Current Assignee / Owner
Priority Date
2025-01-01
Filing Date
2025-08-22
Publication Date
2026-07-09

AI Technical Summary

Technical Problem

Existing nucleic acid capture methods for targeted genome sequencing face challenges such as high cost, low throughput, inefficiency in capturing high GC-rich regions, and PCR bias, leading to poor performance and low yield.

Method used

A method involving single-stranded circular oligonucleotides (circ-oligos) hybridizing with target oligonucleotides, followed by exonuclease treatment, bidirectional Rolling Circle Amplification (RCA), and restriction endonuclease digestion to generate biotinylated nucleic acids for target capture.

Benefits of technology

The method achieves efficient, cost-effective, and scalable generation of nucleic acids for targeted genome enrichment, particularly in high GC-rich regions, with high yield and reduced bias, suitable for next-generation sequencing workflows.

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Abstract

The invention relates to a method and a kit for generating nucleic acid for target capture. The method involves binding of a circ-olio sequence to at least one target oligonucleotide to generate a single stranded circular DNA. The single stranded circular DNA is bidirectionally amplified with the sense and antisense oligonucleotides, followed by biotinylation to obtain for a double-stranded concatemer of linear sequence. The double-stranded concatemer of linear sequence with the restriction endonucleases produces a target capture nucleic acid. The method offers a scalable targeted capture of pr eselected genomic regions, and is cost effective. The method of generating nucleic acid for target capture has application in the field of oncology hematology, inherited malignancies, germline disorders, microbiology and prenatal diagnosis. The invention generates baits over high GC-rich regions of the genome.
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