Abstract: The present invention provides novel engineered polypeptides that support both reverse transcription and DNA amplification in manganese-independent reactions. The present invention also provides methods for amplifying template nucleic acids using such polypeptides. This invention addresses deficiencies in the current state of the art in nucleic acid amplification-based detection of template nucleic acids, especially RNA targets, including deficiencies in detection sensitivity, specificity, enzyme stability, inhibitor tolerance and time to result compared with manganese-dependent thermostable reverse transcriptases and two-enzyme solutions.

Abstract: The present invention relates to a DNA and/or RNA ligase enzyme, its amino acid sequence, its nucleic acid sequence and to DNA and/or RNA ligase proteins encoded by these nucleic acid sequences, as well as nucleic acid or amino acid constructs comprising portions of the nucleic acid or amino acid sequence of the DNA and/or RNA ligase enzyme. The invention further relates to methods of using the DNA and/or RNA ligase enzyme for molecular biological assays and molecular diagnostic applications as well as to a kit containing the DNA and/or RNA ligase enzyme.

Abstract: The present invention provides novel engineered reverse transcriptase enzymes that afford beneficial improvements in thermal stability, processivity, cDNA yields and elimination of secondary enzymatic activity. The present invention also provides methods for amplifying template nucleic acids using such reverse transcriptase enzymes. This invention addresses deficiencies in the current state of the art reverse transcriptase enzymes in RNA detection and analysis including deficiencies in detection sensitivity, specificity, side enzyme activities, enzyme stability and synthesis capacity, especially when using template nucleic acids ranging in length, secondary structure and nucleotide content.

Abstract: The present invention relates to a polymerase enzyme with improved ability to incorporate reversibly terminating nucleotides. The enzyme comprising the following mutations in the motif A region (SGS). It relates to a polymerase enzyme according to SEQ ID NO. 1 with mutations in the amino acid sequence positions 409, 410 and 411.

Abstract: The present invention relates to a polymerase enzyme from Pyrococcus furiosus with improved ability to incorporate reversibly terminating nucleotides. The enzyme comprising the following mutations in the motif A region (SGS). It relates to a polymerase enzyme according to SEQ ID NO. 1 or any polymerase that shares at least 70% amino acid sequence identity thereto, comprising a mutation selected from the group of (i) at position 409 of SEQ ID NO. 3: serine (S) (L409S) and/or, (ii) at position 410 of SEQ ID NO. 3: glycine (G) (Y410G) and/or (iii) at position 411 of SEQ ID NO. 3: serine (S) (P411S), wherein the enzyme has little or no 3?-5? exonuclease activity.

Abstract: The present invention provides novel engineered polypeptides that support both reverse transcription and DNA amplification in manganese-independent reactions. The present invention also provides methods for amplifying template nucleic acids using such polypeptides. This invention addresses deficiencies in the current state of the art in nucleic acid amplification-based detection of template nucleic acids, especially RNA targets, including deficiencies in detection sensitivity, specificity, enzyme stability, inhibitor tolerance and time to result compared with manganese-dependent thermostable reverse transcriptases and two-enzyme solutions.

Abstract: Methods of detecting RNA, such as ribosomal RNA (rRNA), messenger RNA (mRNA), and others. The methods include heating a cell comprising RNA in a solution to release the RNA from the cell, reverse transcribing the RNA into DNA with an enzyme, amplifying the DNA with the same enzyme, and detecting the amplified DNA. The heating, reverse-transcribing, and amplifying in at least some of the methods are performed at substantially the same temperature and a substantially constant temperature without adding additional reagents during or between the steps. The methods can be used to detect the presence of one cell type as distinguished from another cell type within a sample or to determine levels of gene expression, each without the need for elaborate extraction protocols.

Abstract: Thermostable viral and microbial polymerases exhibiting a combination of activities selected from proofreading (3?-5?) exonuclease activity, nick translating (5?-3?) nuclease activity, synthetic primer-initiated polymerase activity, nick-initiated polymerase activity, reverse transcriptase activity, strand displacement activity, terminal transferase activity, primase activity, and/or efficient incorporation of chain terminating analogs. Some of the polymerases provided herein include a first motif and a second motif. The first motif preferably has the sequence X1X2X3DX4PX5IELRX6X7X8, wherein X1 is I or V; X4 is F or Y; X8 is G or A; and X2, X3, X5, X6, and X7 are any amino acid. The second motif preferably has the sequence RX9X10X11KSANX12GX13X14YG, wherein X11 is G or A; X12 is F, L, or Y; X13 is L or V; X14 is I or L; and X9 and X10 are any amino acid.