Roche Sequencing Solutions, Inc.

Kapa Biosystems, Inc.

Signature Diagnostics Gmbh

Roche Diagnostics Gmbh

F. Hoffmann-La Roche Ag

Bronwen Miller

Brian Christopher Godwin

Joseph Platzer

Jo-Anne Elizabeth Penkler

Alexander Lovejoy

Janine Mok

Daniel Burgess

Bernd Hinzmann

Martin Ranik

Sophie Beckert

Daniel Klass

Melissa Loyzer

Victoria Constandse

Improved methods and compositions are provided herein for primer extension target enrichment of target polynucleotides.

Primer extension target enrichment

The present disclosure provides a method for enrichment of at least one target nucleic acid in a library of nucleic acids. A first oligonucleotide is hybridized to a target nucleic acid in library of nucleic acids having first and second adapters. The hybridized first oligonucleotide is extended with a first polymerase, thereby producing a first primer extension complex including the target nucleic acid and the extended first oligonucleotide. The first primer extension complex is captured, enriched relative to the library of nucleic acids, and a second oligonucleotide is hybridized to the target nucleic acid. The hybridized second oligonucleotide is extended with a second polymerase, thereby producing a second primer extension complex including the target nucleic acid and the extended second oligonucleotide, and further liberating the extended first oligonucleotide from the first primer extension complex.

Target enrichment by unidirectional dual probe primer extension

Present disclosure provides a method including isolating DNA from a source, thereby providing a composition including the isolated DNA. The isolated DNA has at least first and second target regions, where the length of the second target region is greater than the length of the first target region. The method further includes quantifying a total mass of the isolated DNA, quantifying a first quantification cycle (C) of the first target region and a second Cof the second target region, and calculating a Q-ratio for the isolated DNA by dividing the second Cby the first C. The method further includes determining a value for a quality-mass constant (k), estimating a required input mass by dividing kby the Q-ratio, and preparing the isolated DNA for sequencing if the total mass of the isolated DNA in the composition is equal or greater than the required input mass.

System and method for isolation and qualification of nucleic acids

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Cape Town, South Africa

Bronwen Miller