Patent ID: 9109222
Filing Date: 2015-08-18
Classification: B01J,C12N,C12Q

Abstract:
1. A method of preparing a nucleic acid library in a multi-purpose buffer, comprising: a) combining a nucleic acid sample with a multi-purpose buffer, wherein said multi-purpose buffer comprises deoxyribonucleotide triphosphates (dNTPs) and primers to generate a mixture comprising said nucleic acid sample and said multi-purpose buffer; b) contacting said mixture comprising said nucleic acid sample and said multi-purpose buffer with a plurality of purified enzymes, wherein said plurality of purified enzymes comprise a first enzyme with polymerase activity, a second enzyme with kinase activity, and a third enzyme with phosphatase activity, and wherein said contacting is under conditions such that amplified nucleic acid is generated in said multipurpose buffer; c) mechanically, physically, chemically or enzymatically treating said amplified nucleic acid in said multi-purpose buffer such that sheared amplified nucleic acid is generated in said multi-purpose buffer without purification of said sheared amplified nucleic acid; d) treating said multi-purpose buffer comprising said sheared amplified nucleic acid to inactivate said polymerase activity, said kinase activity, and said phosphatase activity without purification of said sheared amplified nucleic acid; and e) contacting said multi-purpose buffer comprising said sheared amplified nucleic acid and said inactive polymerase, kinase and phosphatase with a ligase and nucleic acid adapters without purification of said sheared amplified nucleic acid under conditions such that an adapter-linked nucleic acid library is generated.