METHODS FOR NUCLEIC ACID ASSEMBLY AND HIGH THROUGHPUT SEQUENCING

• Main Authors: HUDSON, Michael, E, KUNG, Li-yun A, SCHINDLER, Daniel, ARCHER, Stephen, SAAEM, Ishtiaq
• Format: Patent
• Language: English; French; German
• Published: 03.08.2022
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMBINATORIAL CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; LIBRARIES, e.g. CHEMICAL LIBRARIES, IN SILICOLIBRARIES; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

Methods and apparatus of some aspects of the invention relate to the synthesis of high fidelity polynucleotides. In particular, aspects of the invention relate to concurrent enzymatic removal of amplification sequences and ligation of processed oligonucleotides into nucleic acid assemblies. According to some embodiments, the invention provides a method for producing a target nucleic acid having a predefined sequence. In some embodiments, the method comprises the step of providing a plurality of oligonucleotides, wherein each oligonucleotides comprises (i) an internal sequence identical to a different portion of a sequence of a target nucleic acid, (ii) a 5' sequence flanking the 5' end of the internal sequence and a 3' flanking sequence flanking the 3' end of the internal sequence, each of the flanking sequence comprising a primer recognition site for a primer pair and a restriction enzyme recognition site.