Illumina Corporation

Epicentre Technologies Corporation

Haiying Li Grunenwald

Jerome J Jendrisak

Gary A Dahl

Ramesh Vaidyanathan

Scott Kuersten

Nicholas Caruccio

Molly Min He

Amirali Kia

Erin Bomati

Christian Gloeckner

Trina Faye Osothprarop

Darin J Haskins

Joshua Burgess

Anupama Khanna

Daniel Schlingman

Agnes Radek

Gary Dahl

Jerome Jendrisak

The present invention provides methods, compositions and kits for using a transposase and a transposon end for generating extensive fragmentation and 5'-tagging of double-stranded target DNA in vitro, then using a DNA polymerase for generating 5′- and 3′-tagged single-stranded DNA fragments without performing a PCR amplification reaction, wherein the first tag on the 5′-ends exhibits the sequence of the transferred transposon end and optionally, an additional arbitrary sequence, and the second tag on the 3′-ends exhibits a different sequence from the sequence exhibited by the first tag. The method is useful for generating 5′- and 3′-tagged DNA fragments for use in a variety of processes, including processes for metagenomic analysis of DNA in environmental samples, copy number variation (CNV) analysis of DNA, and comparative genomic sequencing (CGS), including massively parallel DNA sequencing (so-called 'next generation sequencing').

Transposon end compositions and methods for modifying nucleic acids

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

The present invention provides methods, compositions and kits for using a transposase and a transposon end for generating extensive fragmentation and 5'-tagging of double-stranded target DNA in vitro, then using a DNA polymerase for generating 5&#x2032;- and 3&#x2032;-tagged single-stranded DNA fragments without performing a PCR amplification reaction, wherein the first tag on the 5&#x2032;-ends exhibits the sequence of the transferred transposon end and optionally, an additional arbitrary sequence, and the second tag on the 3&#x2032;-ends exhibits a different sequence from the sequence exhibited by the first tag. The method is useful for generating 5&#x2032;- and 3&#x2032;-tagged DNA fragments for use in a variety of processes, including processes for metagenomic analysis of DNA in environmental samples, copy number variation (CNV) analysis of DNA, and comparative genomic sequencing (CGS), including massively parallel DNA sequencing so-called 'next generation sequencing').

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 invention provides methods, compositions and kits for using a transposase and a transposon end for generating extensive fragmentation and 5'-tagging of double-stranded target DNA in vitro, then using a DNA polymerase for generating 5&#x2032;- and 3&#x2032;-tagged single-stranded DNA fragments without performing a PCR amplification reaction, wherein the first tag on the 5&#x2032;-ends exhibits the sequence of the transferred transposon end and optionally, an additional arbitrary sequence, and the second tag on the 3&#x2032;-ends exhibits a different sequence from the sequence exhibited by the first tag. The method is useful for generating 5&#x2032;- and 3&#x2032;-tagged DNA fragments for use in a variety of processes, including processes for metagenomic analysis of DNA in environmental samples, copy number variation (CNV) analysis of DNA, and comparative genomic sequencing (CGS), including massively parallel DNA sequencing (so-called 'next generation sequencing).

Compositions of transposome complexes for generating DNA fragments with specific 5'- and 3&#x2032;-tags. Kits for generating libraries for sequencing, with transposome complexes, enzymes, oligonucleotides or other components.

The present invention provides methods, compositions and kits for using a transposase and a transposon end for generating extensive fragmentation and 5'-tagging of double-stranded target DNA in vitro, then using a DNA polymerase for generating 5&#x2032;- and 3&#x2032;-tagged single-stranded DNA fragments without performing a PCR amplification reaction, wherein the first tag on the 5&#x2032;-ends exhibits the sequence of the transferred transposon end and optionally, an additional arbitrary sequence, and the second tag on the 3&#x2032;-ends exhibits a different sequence from the sequence exhibited by the first tag. The method is useful for generating 5&#x2032;- and 3&#x2032;-tagged DNA fragments for use in a variety of processes, including processes for metagenomic analysis of DNA in environmental samples, copy number variation (CNV) analysis of DNA, and comparative genomic sequencing (CGS), including massively parallel DNA sequencing (so-called 'next-generation sequencing.).

The present invention provides methods, compositions and kits for using a transposase and a transposon end for generating extensive fragmentation and 5'-tagging of double-stranded target DNA in vitro, then using a DNA polymerase for generating 5&#x2032;- and 3&#x2032;-tagged single-stranded DNA fragments without performing a PCR amplification reaction, wherein the first tag on the 5&#x2032;-ends exhibits the sequence of the transferred transposon end and optionally, an additional arbitrary sequence, and the second tag on the 3&#x2032;-ends exhibits a different sequence from the sequence exhibited by the first tag. The method is useful for generating 5&#x2032;- and 3&#x2032;-tagged DNA fragments for use in a variety of processes, including processes for metagenomic analysis of DNA in environmental samples, copy number variation (CNV) analysis of DNA, and comparative genomic sequencing (CGS), including massively parallel DNA sequencing (so-called 'next-generation sequencing.)

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Belleville, WI, United States of America

Haiying Li Grunenwald