Abstract:
The present invention provides methods and compositions for improving the efficiency of nucleic acid amplification reactions. The invention encompasses hybrid polymerases that show increased processivity over wild type polymerases as well as decreased exonucleases activity. The invention also encompasses methods, compositions and kits for conducting nucleic acid synthesis and amplification reactions in which non-specific amplification of primers is reduced.
Abstract:
Comparison of common sequencing reads from sequencing based on partition-based barcoding can be used to improve sequencing results. Increased loading of barcodes per partition can also improve sequencing results.
Abstract:
The present invention provides methods and compositions for improving the efficiency of nucleic acid amplification reactions. The invention encompasses hybrid polymerases that show increased processivity over wild type polymerases as well as decreased exonucleases activity. The invention also encompasses methods, compositions and kits for conducting nucleic acid synthesis and amplification reactions in which non-specific amplification of primers is reduced.
Abstract:
The present invention provides methods for improving the specificity of nucleic acid amplification comprising incubating a nucleic acid molecule with a polymerase-Sso7 DNA binding domain conjugate and arginine, spermidine, or spermine. The present invention also provides reaction mixtures and kits for improving the specificity of nucleic acid amplification.
Abstract:
Methods of determining polymerase fidelity are provided. In one embodiment, the method comprises filling a gapped plasmid with a polymerase to form a gap-filled plasmid, wherein the gap-filled plasmid comprises a gene encoding an protein that is functional or non-functional depending on the polymerase fidelity; forming a plurality of partitions from a solution comprising the gap-filled plasmid and a label for detecting the presence of the plasmid; detecting the presence of the gap-filled plasmid in one or more of the partitions; and determining the fidelity of the polymerase by determining a ratio of partitions containing the gene encoding a functional protein to partitions containing a gene encoding a non-functional protein.
Abstract:
Fusion polypeptides having a heterologous 5′-3′ exonuclease domain linked to a polymerase that does not naturally have 5′-3′ exonuclease activity, as well as methods of their use are provided. Other aspects are also disclosed.