Siemens Healthcare Diagnostics Gmbh

Thomas Battersby

Adrian Nielsen Fehr

Minxue Zheng

Toumy Guettouche

James Hnatyszyn

Stacey Tam

Maria Albalos

Edward Brooks

Mark Baumeister

Michel Friesenhahn

Jesse Brooks

Felix Kleshik

The present disclosure relates to normalization of biological samples, particularly samples comprising nucleic acids to be sequenced. The normalization protocols described herein may be utilized across multiple samples to cap total stoichiometric input and minimize variations in transcript abundance on a per-sample basis in a multiplexed fashion to dramatically increase the accuracy, capacity and efficiency of nucleic acid sequencing.

Method of normalizing biological samples

The present invention relates to amplification primers with a universal tag and sequencing primers comprising at least one non-natural nucleobase capable of hybridizing to a complementary non-natural nucleobase. The present invention further relates to amplification methods of nucleic acid amplification and sequencing using an amplification primer with a universal tag and sequencing primers, as well as kits and solid supports comprising such primers and tags.

Universal tags with non-natural nucleobases

The present invention relates to novel degenerate nucleobase analogs and degenerate nucleobase oligomers derived therefrom, and methods of using such degenerate nucleobase oligomers.

Degenerate nucleobase analogs

The methods and reagents described herein can be used to shorten incubation times in hybridization assays. As demonstrated in the examples, we have identified specific sulfonic acid polymers and hybridization conditions that lead to significantly shorter incubation times (e.g., signals after three hours that are comparable to signals that could traditionally only be obtained after overnight incubation). In some embodiments, shorter incubation times are achieved by adding the sulfonic acid polymer(s) during the hybridization process. Alternatively or additionally, in some embodiments, shorter incubation times are achieved via changes to the hybridizing conditions, e.g., by reducing the hybridization volume, increasing the salt concentration, andor increasing the probe concentration (capture extender probe andor label extender probe).

Methods and reagents for shortening incubation times in hybridization assays

The present invention relates to stable anti-parallel heteropolynucleotide duplexes, comprising a plurality of complementary purine-purine nucleobase dyads, wherein the nucleobase is coupled to a pentose sugar backbone. The present invention further relates to methods of hybridizing two heteropolynucleotide molecules to form such purine-purine nucleobase dyads, as well as kits and solid supports comprising such purine-purine nucleobase dyads.

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El Cerrito, CA, United States of America

Thomas Battersby