Invention Grant
- Patent Title: Method to screen for a mutant within a population of organisms by applying a pooling and splitting approach
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Application No.: US16311450Application Date: 2017-06-23
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Publication No.: US11884972B2Publication Date: 2024-01-30
- Inventor: Toni Wendt , Ole Olsen , Søren Knudsen , Hanne Cecille Thomsen , Birgitte Skadhauge , Magnus Wohlfahrt Rasmussen , Massimiliano Carciofi , Alexander Striebeck
- Applicant: Carlsberg A/S
- Applicant Address: DK Copenhagen
- Assignee: Carlsberg A/S
- Current Assignee: Carlsberg A/S
- Current Assignee Address: DK Copenhagen V
- Agency: McNeill Baur PLLC
- Priority: DK 201670485 2016.07.01
- International Application: PCT/EP2017/065516 2017.06.23
- International Announcement: WO2018/001884A 2018.01.04
- Date entered country: 2018-12-19
- Main IPC: C12Q1/68
- IPC: C12Q1/68 ; C12Q1/686 ; C12N15/01 ; C12N15/10 ; A01H6/46 ; C12Q1/6806 ; C12Q1/6827 ; C12Q1/6876 ; C40B30/00 ; C40B40/02

Abstract:
In traditional plant breeding approaches, chemical mutagenesis may be utilized to introduce nucleotide substitutions at random in the genome of a plant, i.e. without possibilities to control the sites of nucleotide changes. Because of genome complexities, the statistical probability is extremely little when it comes to finding a predetermined nucleotide substitution. The present invention, however, demonstrates how a novel, alternative use of digital polymerase chain reaction (dPCR), preferably droplet dPCR (ddPCR), is developed to exploit finding of specific nucleotide substitutions in mutated genes. The entire platform comprises a screening method with a library of mutagenized organisms, digital PCR-based systems and a set-up to propagate and analyze identified, mutated organisms.
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