Thermo Fisher Scientific Baltics Uab

Fermentas Uab

Life Technologies Corporation

Vilnius University

Fermentas Ab

Arvydas Lubys

Arvydas Janulaitis

Renata Rimseliene

Mindaugas Ukanis

Judita Lubiene

Zana Kapustina

Arturas Berezniakovas

Ervinas Gaidamauskas

Romas Tamosevicius

Mark R Andersen

Virginijus Siksnys

Sihong Chen

Lolita Zaliauskiene

Michael D Allen

Inga Cikotiene

Giedrius Gasiunas

Jolanta Vitkuté

Remigijus Skirgalia

Aiste Jaspone

Justina Medziune

Simona Zeimyte

Anja Smith

Monika Gasiuniene

Algimantas Markauskas

Komelijus Stankevicius

Monika Glemzaite

This application relates to mutant polymerases. This application discloses mutant phage-type RNA polymerases, such as a mutant T7, SP6, and T3 RNA polymerase, may use 2'-modified nucleoside triphosphates or deoxynucleotide triphosphates as substrates. Methods for producing nucleic acid molecules using these mutant polymerases are also disclosed.

Mutant polymerases and methods of using the same

The present disclosure describes oligonucleotide-tethered nucleotides, methods of making them, and methods of using them. The oligonucleotide-tethered nucleotides comprise, in some embodiments, a nucleotide linked to an oligonucleotide of from about 3 to about 100 nucleotides in length. These oligonucleotide-tethered nucleotides can be used to label a plurality of different types of nucleic acids in a plurality of different situations with a known oligonucleotide, which can serve as a barcode in some embodiments. The resulting oligonucleotide-labeled nucleic acids oligonucleotides can be used in a variety of nucleic acid sequencing methods.

Oligonucleotide-tethered nucleotides

Isolation or in vitro assembly of the Cas9-crRNA complex of theCRISPR3/Cas system and use for cleavage of DNA bearing a nucleotide sequence complementary to the crRNA and a proto-spacer adjacent motif. Methods for site-specific modification of a target DNA molecule in vitro or in vivo using an RNA-guided DNA endonuclease comprising RNA sequences and at least one of an RuvC active site motif and an HNH active site motif; for conversion of Cas9 polypeptide into a nickase cleaving one strand of double-stranded DNA by inactivating one of the active sites (RuvC or HNH) in the polypeptide by at least one point mutation; for assembly of active polypeptide-polyribonucleotides complex in vivo or in vitro; and for re-programming a Cas9-crRNA complex specificity in vitro and using a cassette containing a single repeat-spacer-repeat unit.

RNA-directed DNA cleavage by the Cas9-crRNA complex

A composition and method for controlled in vitro fragmentation of nucleic acids. A transposase forms catalytically active complexes with a modified transposon end that contains within its end sequence degenerate, apurinic/apyrimidinic sites, nicks, or nucleotide gaps, to fragment or shear a target nucleic acid sample in a controlled process. This method yields desired average nucleic acid fragment sizes. The inventive composition and method may be applied for generation of DNA fragments containing shortened transposon end sequences to facilitate subsequent reactions, for production of asymmetrically tailed DNA fragments, etc.

Method for controlled DNA fragmentation

Enzyme compositions and their method of use that provide ready-to-use master mixtures. The compositions comprise a modified thermophilic DNA polymerase lacking 5'-3&#x2032; and 3&#x2032;-5&#x2032; exonuclease activity premixed with T4 DNA polymerase, Klenow fragment and T4 polynucleotide kinase and all other necessary components, including reaction buffer and nucleoside triphosphates, required to perform DNA blunting, phosphorylation, and single nucleotide extension reactions in one tube and in two steps. Among other benefits, the mixture of different enzymes, buffers and nucleoside triphosphates is stable during prolonged storage.

Enzyme composition for DNA end repair, adenylation, phosphorylation

A vector for transformation into a host cell is described comprising a toxic gene encoding a product that is lethal to the host cell, wherein the toxic gene comprises:

Vectors for cloning

A restriction endonuclease with a recognition sequence 5'-TCGA-3&#x2032;. The restriction endonuclease is sensitive to the presence of a modified cytosine residue in the recognition sequence. Methods and kits using the restriction endonuclease with a recognition sequence 5&#x2032;-TCGA-3&#x2032; are also disclosed.

Restriction endonucleases and their uses

Provided is a methylation-specific restriction endonuclease for a DNA duplex substrate, which endonuclease recognizes in a strand of the duplex a 2 to 6 nucleotide recognition sequence comprising a 5-methylcytosine, and cleaves each strand of the duplex at a fixed position outside the recognition sequence.

Restriction endonucleases and their applications

A process for producing a polynucleotide encoding a restriction endonuclease with an altered specificity, which process comprises:

Nuclease

A strand-specific polynucleotide nickase comprising an endonuclease which comprises a first subunit and a second subunit and which recognises an asymmetric nucleotide recognition sequence, wherein the first subunit comprises a catalytic domain capable of cleaving one strand of a DNA duplex, and the second subunit is incapable of cleaving the other strand of the DNA duplex.

Strand-specific polynucleotide nickases

Growing community of inventors

Vilnius, Lithuania

Arvydas Lubys