Methods for preparing a next generation sequencing (NGS) library from a ribonucleic acid (RNA) sample and compositions for practicing the same

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United States of America Patent

PATENT NO 11959078
APP PUB NO 20210047638A1
SERIAL NO

17076568

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Abstract

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Methods of preparing a next generation sequencing (NGS) library from a ribonucleic acid (RNA) sample are provided. Aspects of the methods include combining the RNA sample with a first strand cDNA primer and a template switch oligonucleotide under first strand cDNA synthesis conditions, where one of the first strand cDNA primer and the template switch oligonucleotide includes a first post-tagmentation amplification primer binding domain. The resultant product is subjected to amplification conditions sufficient to produce a double stranded cDNA, which is then tagmented with a transposome that includes a second post-tagmentation amplification primer binding domain. The tagmented sample is then subjected to amplification conditions using first and second post-tagmentation amplification primers that include sequencing platform adapter constructs to produce a NGS library. Aspects of the invention further include compositions produced by the methods and kits that find use in practicing the methods.

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Patent Owner(s)

  • TAKARA BIO USA, INC.

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Inventor(s)

Inventor Name Address # of filed Patents Total Citations
Bostick, Magnolia San Mateo, US 11 11
Chang, Cynthia San Mateo, US 3 100

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