Method to Screen For a Mutant Within a Population of Organisms By Applying a Pooling and Splitting Approach
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United States of America Patent
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Issued Date -
Jun 6, 2024
app pub date -
Dec 12, 2023
filing date -
Jul 1, 2016
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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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