Patent application number | Description | Published |
20110124055 | Methods for High Fidelity Production of Long Nucleic Acid Molecules - In a method for synthesizing a long nucleic acid molecule, a first immobilized nucleic acid has a first 5′ region and a first 3′ region and a second immobilized nucleic acid has a second 5′ region and a second 3′ region, wherein the second 3′ region and the first 5′ region have identical nucleic acid sequences. The first immobilized nucleic acid is hybridized with an oligonucleotide under conditions promoting hybridization of the oligonucleotide to the first 3′ region, extending the hybridized oligonucleotide and producing a first extension product having a 3′ region that is complementary to the first 5′ region. The second immobilized nucleic acid is hybridized with the first extension product under conditions promoting hybridization of the 3′ region of the first extension product to the second 3′ region, extending the 3′ region of the first extension product and producing a second extension product having a 3′ region that is complementary to the second 5′ region, wherein the second extension product has a sequence complementary to the first and second 3′ and 5′ regions. | 05-26-2011 |
20110201057 | Methods for High Fidelity Production of Long Nucleic Acid Molecules with Error Control - A method for synthesizing a nucleic acid having a desired sequence and length comprises providing a solid support having an immobilized nucleic acid, performing a nucleic acid addition reaction to elongate the immobilized nucleic acid by adding a nucleotide or an oligonucleotide to the nucleic acid, determining whether the nucleotide or the oligonucleotide is added to the nucleic acid by detecting whether there is an increase in electrophoretic force applied to the solid support when an electric field and a magnetic field gradient are applied to the support, wherein the increase in electrophoretic force applied to the support is caused by adding the nucleotide or the oligonucleotide to the nucleic acid, repeating the addition reaction and determination steps if the nucleotide or the oligonucleotide is not added to the nucleic acid, and continuing until the immobilized nucleic acid has a desired sequence and length. | 08-18-2011 |
20120264653 | Methods for High Fidelity Production of Long Nucleic Acid Molecules - In a method for synthesizing a pool of nucleic acid molecules, a first nucleic acid has a first 5′ region and a first 3′ region and a second nucleic acid has a second 5′ region and a second 3′ region. The second 3′ region and the first 5′ region have identical nucleic acid sequences. The first 3′ region is hybridized with an oligonucleotide, extending the hybridized oligonucleotide and producing a first extension product having a 3′ region complementary to the first 5′ region. The second nucleic acid is hybridized with the first extension product to hybridize the 3′ region of the first extension product to the second 3′ region, extending the 3′ region of the first extension product and producing a second extension product having a 3′ region complementary to the second 5′ region. Error-containing molecules are separated from error-free molecules by a component that selects for a sequence error. | 10-18-2012 |
20130005612 | Methods for High Fidelity Production of Long Nucleic Acid Molecules with Error Control - A method for synthesizing a nucleic acid having a desired sequence and length comprises providing a solid support having an immobilized nucleic acid, performing a nucleic acid addition reaction to elongate the immobilized nucleic acid by adding a nucleotide or an oligonucleotide attached to a protecting group to the nucleic acid, determining whether the nucleotide or the oligonucleotide is added to the nucleic acid, removing the protecting group, and continuing until the immobilized nucleic acid has a desired sequence and length. | 01-03-2013 |
20130323722 | Methods for High Fidelity Production of Long Nucleic Acid Molecules - In a method for generating a long nucleic acid molecule, nucleic acids immobilized on a surface and having overlapping complementary sequences is released into solution. The overlapping complementary sequences are hybridized to form hybridized nucleic acids, followed by extension or ligation of the hybridized nucleic acids to synthesize the long nucleic acid molecule. The nucleic acids may comprise first and second series of nucleic acids having redundant overlapping sequences, wherein nucleic acids from the first and second series are complementary to each other. The complementary nucleic acids are hybridized to form the hybridized nucleic acids. The generated long nucleic acid molecule may have a predetermined sequence element, and it may be introduced into a system wherein the predetermined sequence element is required for replication, such that replication of the synthesized long nucleic acid molecule is indicative of the presence of the predetermined sequence element in the long nucleic acid molecule. | 12-05-2013 |
20140349400 | Programmable Modification of DNA - A self-reconfiguring genome uses a cassette having operons or DNA sequences that code for guide RNA, reverse transcriptase, donor RNA, and a CRISPR cleavage enzyme. A self-reconfiguring genome may be based on lambda recombineering of in situ generated oligonucleotides. A method for programmable self-modification of a cellular genome includes transcribing guide RNA from a self-reconfiguring cassette, associating the transcribed guideRNA with the CRISPR enzyme, intercalcating a region of complimentary sequence within an integration site of the genome, cutting upstream of a PAM site within the integration site; transcribing the donorRNA, translating donorRNA to double-stranded DNA, and recombining the double-stranded DNA via homologous recombination at the cut site of the integration site. A set of cascadable and multiplexable genetic logic gates with a universal RNA input/output based on single-strand annealing or non-homologous end joining, comprises transcription promoters or terminators, homologous regions, DNA sequences, RNA, and enzymes from the CRISPR system. | 11-27-2014 |
20150064791 | Microfluidic-based Gene Synthesis - A method for synthesizing long DNA constructs from oligonucleotide precursors directly within a microfluidic device uses several oligonucleotides at once. A precursor mix containing at least two oligonucleotide precursors with at least partial base complementarity is introduced into an input of a microfluidic chip and at least one cycle of at least one gene synthesis protocol is applied to fabricate a DNA construct containing the sequence of at least two oligonucleotide precursors. A method for the synthesis of a modified DNA construct includes electroporating at least one oligonucleotide encoding for at least one point mutation and having homology with at least one DNA region of a target cell into the target cell and incorporating the oligonucleotide into the target cell DNA through the action of recombination protein beta or a recombination protein beta functional homolog. | 03-05-2015 |
20150283514 | NANOFLUIDIC SORTING SYSTEM FOR GENE SYNTHESIS AND PCR REACTION PRODUCTS - Devices and methods integrate nanopore and microfluidic technologies for recording molecular characteristics of individual molecules such as, for example, biomolecules. Devices comprise a first substrate comprising a microchannel, a second substrate comprising a microchannel, the second substrate positioned below the first substrate, and a membrane having a thickness of about 0.3 nm to about 1 nm and comprising at least one nanopore, the membrane positioned between the first substrate and the second substrate, wherein a single nanopore of the membrane is constructed and arranged for electrical and fluid communication between the microchannel of the first substrate and the microchannel of the second substrate. To mitigate the effect of errors that occur during de novo DNA synthesis, longer DNA molecules are typically synthesized from shorter oligonucleotides by polymerase construction and amplification (PCA), or by other methods. | 10-08-2015 |