Illumina Corporation

Epicentre Technologies Corporation

University of Texas System

Scott Kuersten

Ramesh Vaidyanathan

Haiying Li Grunenwald

Molly Min He

Amirali Kia

Erin Bomati

Christian Gloeckner

Darin J Haskins

Trina Faye Osothprarop

Joshua Burgess

Anupama Khanna

Daniel Schlingman

Agnes Radek

Ken Doyle

Frederick W Hyde

Asako Tetsubayashi

Alan M Lambowitz

Sabine Mohr

Travis B White

The present disclosure is related to methods and materials for depleting unwanted RNA species from a nucleic acid sample. In particular, the present disclosure describes how to remove unwanted rRNA, tRNA, mRNA or other RNA species that could interfere with the analysis, manipulation and study of target RNA molecules in a sample.

Nuclease-based RNA depletion

Presented herein are methods and compositions for analyzing rare nucleic acid species. Some methods presented herein use DNA reassociation kinetics following thermal denaturation to define populations of nucleic acid sequences, e.g., highly abundant (e.g., cDNA from rRNA), moderately abundant, and less abundant or rare sequences (e.g., cDNA from mRNA).

Methods of analyzing nucleic acids

Presented herein are transposase enzymes and reaction conditions for improved fragmentation and tagging of nucleic acid samples, in particular altered transposases and reaction conditions which exhibit improved insertion sequence bias, as well as methods and kits using the same.

Modified transposases for improved insertion sequence bias and increased DNA input tolerance

The present innovation provides methods and kits that enable rapid and efficient dual end-tagging of RNA to prepare libraries for analysis by applications such as next-generation RNA sequencing, qPCR, microarray analysis, or cloning. The methods do not require time-consuming and inefficient gel-purification steps that are common to methods known in the art. In addition, the present invention provides methods and kits for rapid, high-throughput enzymatic preparation of 5'-activated, 3&#x2032;-blocked DNA oligonucleotides from standard, single-stranded DNA oligonucleotides.

Methods and kits for 3'-end-tagging of RNA

A method of preparing a DNA copy of a target polynucleotide using template switching is described. The method includes mixing a double stranded template/primer substrate made up of a DNA primer oligonucleotide associated with a complementary oligonucleotide template strand with a target polynucleotide in a reaction medium and adding a suitable amount of a non-retroviral reverse transcriptase to the reaction medium to extend the DNA primer oligonucleotide from its 3' end to provide a DNA copy polynucleotide. The DNA copy polynucleotide includes a complementary target DNA polynucleotide that is synthesized using the target polynucleotide as a template. Methods of adding nucleotides to the double stranded template/primer substrate are also described. The method can be used to facilitate detection, PCR amplification, cloning, and determination of RNA and DNA sequences.

Use of template switching for DNA synthesis

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Madison, WI, United States of America

Scott Kuersten