The invention relates to methods for amplifying a DNA template, comprising incubating the DNA template with a DNA-dependent RNA polymerase in the presence of ribonucleotides and amplifying the DNA template by a strand-displacing DNA polymerase. The invention further relates to the use of a RNA polymerase for generating a ribonucleotide primer on a DNA template, followed by amplification of the DNA template by a strand-displacing DNA polymerase, to a kit of parts, comprising a RNA polymerase and a strand-displacing DNA-dependent DNA polymerase, and to the use of the kit of parts for amplification of a DNA template.

The invention relates to methods for amplifying a DNA template, comprising incubating the DNA template with a DNA-dependent RNA polymerase in the presence of ribonucleotides and amplifying the DNA template by a strand-displacing DNA polymerase. The invention further relates to the use of a RNA polymerase for generating a ribonucleotide primer on a DNA template, followed by amplification of the DNA template by a strand-displacing DNA polymerase, to a kit of parts, comprising a RNA polymerase and a strand-displacing DNA-dependent DNA polymerase, and to the use of the kit of parts for amplification of a DNA template.

Disclosed herein are shelf-stable compositions based on synthetic gene networks and/or cell-free systems that are lyophilized on a solid support. The compositions can be easily transported and stored for a period of time, and activation can be done by simply adding water. Methods of use are also disclosed herein, including, but are not limited to, sensing and a variety of logic functions. The invention permits straightforward, sterile and abiotic distribution of synthetic biology-based technology to clinical settings, food processing and industry, the military and consumer products.

Disclosed herein are shelf-stable compositions based on synthetic gene networks and/or cell-free systems that are lyophilized on a solid support. The compositions can be easily transported and stored for a period of time, and activation can be done by simply adding water. Methods of use are also disclosed herein, including, but are not limited to, sensing and a variety of logic functions. The invention permits straightforward, sterile and abiotic distribution of synthetic biology-based technology to clinical settings, food processing and industry, the military and consumer products.

Provided herein, in some aspects, are methods and compositions for cell-free production of ribonucleic acid.

Provided herein, in some aspects, are methods and compositions for cell-free production of ribonucleic acid.

Described herein are synthetic methods for producing sequence-specific RNA oligonucleotides that eliminate impurities produced in prior art methods. In one aspect, a first amplification primer includes one or more deoxyuridine residues, wherein at least one of the one or more deoxyuridine residues is at position −1, −2, −3, −4 or −5 of the promoter region for the single-subunit, DNA-dependent RNA polymerase. The deoxyuridines are excised to provide an amplified functional template DNA which is then used to synthesize RNA which has reduced immunogenic double stranded RNA compared to controls.

Described herein are synthetic methods for producing sequence-specific RNA oligonucleotides that eliminate impurities produced in prior art methods. In one aspect, a first amplification primer includes one or more deoxyuridine residues, wherein at least one of the one or more deoxyuridine residues is at position −1, −2, −3, −4 or −5 of the promoter region for the single-subunit, DNA-dependent RNA polymerase. The deoxyuridines are excised to provide an amplified functional template DNA which is then used to synthesize RNA which has reduced immunogenic double stranded RNA compared to controls.

The disclosure relates to methods of preparing single-stranded DNA (ssDNA). The ssDNA can be used, for example, to prepare functionalized alignment beads as fiducial markers to improve image registration in fluorescence assays for the detection and quantitation of analytes in a sample.

The disclosure relates to methods of preparing single-stranded DNA (ssDNA). The ssDNA can be used, for example, to prepare functionalized alignment beads as fiducial markers to improve image registration in fluorescence assays for the detection and quantitation of analytes in a sample.