Disclosed herein are modified polymerase compositions exhibiting altered polymerase activity, which can be useful in a variety of biological applications. Also disclosed herein are methods of making and using such compositions. In some embodiments, the compositions exhibit altered properties that can enhance their utility in a variety of biological applications. Such altered properties, can include, for example, altered nucleotide binding affinities, altered nucleotide incorporation kinetics, altered photostability and/or altered nanoparticle tolerance, as well as a range of other properties as disclosed herein.

Compositions comprising modified recombinant polymerizing enzymes are provided, along with nucleic acid molecules encoding the modified polymerizing enzymes. In some aspects, methods of using such polymerizing enzymes to synthesize a nucleic acid molecule or to sequence a nucleic acid template are provided.

Provided are HBV immunogenic polypeptides, polynucleotides encoding such polypeptides, vectors expressing such immunogenic polypeptides for use in eliciting an immune response against HBV; pharmaceutical and immunogenic compositions and kits comprising such polypeptides, polynucleotides or vectors, and methods of use in treating and/or preventing HBV.

The present disclosure includes compositions and methods for improved DNA amplification reactions. In particular, the present disclosure provides compositions and methods for hot-start PCR applications using DNA polymerase inhibitors that minimize non-specific DNA amplification by inactivating DNA polymerase at lower temperatures.

Provided herein are compositions and methods using mutant Phi29 polymerases for nucleic acid amplification. Further provided herein are methods for accurate and scalable Primary Template-Directed Amplification (PTA) nucleic acid amplification and sequencing methods, and their applications for mutational analysis in research, diagnostics, and treatment using mutant Phi29 polymerases.

The invention includes a mutant Taq polymerase, which can effectively amplify a target sequence under conditions of salt concentration(s) similar to body fluids, including blood, serum or plasma preserved with sodium citrate. The mutant Taq polymerase, or a biologically active fragment thereof, has one or more substitutions differing from the wild type as shown in Table I.

The present invention provides a DNA polymerase including the sequence of SEQ ID NO. 1 or a sequence which is at least 70% identical thereto, but wherein the aspartic acid residue at position 18 of SEQ ID NO. 1, or the equivalent aspartic acid residue in other sequences, has been replaced by a non-negatively charged amino acid residue. It further provides DNA polymerases comprising the amino acid sequences of SEQ ID NO. 2, 11 and 12 and variants thereof. The present invention also provides nucleic acids encoding the DNA polymerases, a method of producing said DNA polymerases, and compositions, expression vectors and host cells or viruses comprising said DNA polymerases. The present invention also provides uses of said DNA polymerases in nucleotide polymerisation, amplification, and sequencing reactions.

An apparatus for preparing nucleic acid sequences using an enzyme, including a reactor, a plurality of nucleotide material bottles, a deblocking material bottle, and a liquid delivering device. The reactor includes a reaction substrate having a pretreated surface. Each of the nucleotide material bottles is adapted to contain a first reaction solution, and the first reaction solution includes a reaction enzyme and a nucleotide having a terminal protecting group. The deblocking material bottle is adapted to contain a deblocking solution. The liquid delivering device is connected to the reactor, the nucleotide material bottles and the deblocking material bottle. The reaction enzyme is adapted to dispose the nucleotide having the terminal protecting group on the pretreated surface. The reactor has an operating temperature of 45° C.-105° C. A method for preparing nucleic acid sequences using an enzyme of the invention is provided.

Methods of amplifying an RNA template according to aspects of the present disclosure include: providing a composition including a recombinant thermostable DNA polymerase including SEQ ID NO:1, or a variant thereof having at least 99% identity to SEQ ID NO:1; and a reaction buffer, the reaction buffer including 10-30 mM Tris-HCl, pH 8.5-9.0; 20-40 mM KCl; 5-15 mM (NH.sub.4).sub.2SO.sub.4; 1.5-3.5 mM Mn.sup.2+; 0.01-0.20% (w/v) gelatin and/or serum albumin; 0.05-0.15% (w/v) of a nonionic detergent; with the proviso that no more than 0.1 mM of Mg.sup.2+ is present in the composition; and, optionally, 0.01-0.1 mM of a chelating agent is present in the composition.

The invention includes a mutant Taq polymerase or biologically active fragment thereof, which can significantly extend and amplify a target sequence in comparison to wild type Taq polymerase, where the extension mixture includes asymmetrical cyanine dye, including SYBR Green I. The mutant Taq polymerase, or a biologically active fragment thereof, has one or more substitutions differing from the wild type as shown in Table I.