Provided is a linker element and a method of using the linker element to construct a sequencing library, wherein the linker element consists of a linker A and a linker B, the linker A is obtained through the complementary pairing of a long nucleic acid strand and a short nucleic acid strand, the 5′ end of the long strand has a phosphoric acid modification, and the 3′ end of the short strand has an enclosed modification, with enzyme sites in the short strand; and the linker B is a nucleic acid single strand, and the 3′ end thereof can be in a complementary pairing with the 5′ end of the long strand of the linker A. Using the linker element of the present invention for constructing a sequencing library ensures the linking directionality of the linkers while solving the problems of fragment interlinking, linker self-linking and low linking efficiency, and reducing the purification reaction between steps, shortening the linking time and reducing costs.

The disclosure provides a composition comprising a double-stranded deoxyribonucleic acid (dsDNA) sequence comprising from 5′ to 3′, a sequence comprising a first adaptor sequence, a template sequence, and a second adaptor sequence, wherein the second adaptor sequence comprises a hybridization site for a template switching oligonucleotide (TSO). The disclosure provides methods for making the compositions of the disclosure using a template switching mechanism to add non-templated basepairs to the ends of a DNA molecule, hybridize a TSO to the non-templated basepairs, and then extend the sequence complementary to the TSO to add an adaptor.

Described herein are stabilized polymerase compositions comprising a polymerase and an polymerase stabilizing agent, such as a non-detergent zwitterionic stabilizer or a cationic ester disclosed, for use in nucleic acid amplification or nucleic acid sequencing. Compositions are provided for the stabilization of one or more polymerases in a single stabilized liquid formulation. Also disclosed are methods for making and using stabilized polymerase compositions and kits for nucleic acid amplification and sequencing comprising the stabilized polymerase compositions provided.

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.

The invention provides a more efficient and less error-prone method of performing LAMP. The invention also provides a method for utilizing an RNase H2-cleavable probe as a technique for generating signal from the reaction, potentially increasing the specificity of the signal generation.

The present disclosure provides methods and compositions for enhancing the processivity of a polymerase in catalyzing template-dependent DNA synthesis in high concentrations of salt. Also disclosed are methods and compositions for enhancing the assembly of polymerase-template complex compatible with active DNA synthesis in the presence of low levels of nucleotides and at a high temperature, such as temperatures at or near the melting temperature of the polymerase.

Described herein are polymerase variants that are exonuclease deficient. Some variants retain the strand displacement capability comparable to the wild-type or parental polymerase. Some variants have a strand displacement capability that is improved relative to the wild-type or parental polymerase. The variants may have an extension rate that is greater than the wild-type or parental polymerase. The variants may have a waiting time that is less than the wild-type or parental polymerase.

The present invention provides a method of amplifying an RNA molecule in a biological sample by reverse transcription PCR (RT-PCR), wherein the RT-PCR is carried out in a solution comprising a polar aprotic solvent; a serum albumin, and a polyol.

Disclosed are mutant DNA polymerases having increased 3′-mismatch discrimination relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.

Disclosed are mutant DNA polymerases having increased 3′-mismatch discrimination relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.