An example method includes reacting a first solution and a different, second solution on a flow cell by flowing the first solution over amplification sites on the flow cell and subsequently flowing the second solution over the amplification sites. The first solution includes target nucleic acids and a first reagent mixture that comprises nucleoside triphosphates and replication enzymes. The target nucleic acids in the first solution transport to and bind to the amplification sites at a transport rate. The first reagent mixture amplifies the target nucleic acids that are bound to the amplification sites to produce clonal populations of amplicons originating from corresponding target nucleic acids. The amplicons are produced at an amplification rate that exceeds the transport rate. The second solution includes a second reagent mixture and lacks the target nucleic acids. The second solution is to increase a number of the amplicons at the amplification sites.

An example method includes reacting a first solution and a different, second solution on a flow cell by flowing the first solution over amplification sites on the flow cell and subsequently flowing the second solution over the amplification sites. The first solution includes target nucleic acids and a first reagent mixture that comprises nucleoside triphosphates and replication enzymes. The target nucleic acids in the first solution transport to and bind to the amplification sites at a transport rate. The first reagent mixture amplifies the target nucleic acids that are bound to the amplification sites to produce clonal populations of amplicons originating from corresponding target nucleic acids. The amplicons are produced at an amplification rate that exceeds the transport rate. The second solution includes a second reagent mixture and lacks the target nucleic acids. The second solution is to increase a number of the amplicons at the amplification sites.

The present invention relates to an improved process for synthesis of deoxyribonucleic acid (DNA), in particular cell-free enzymatic synthesis of DNA, preferably on a large or industrial scale, with an improved yield and/or with an improved efficiency. The invention requires the use of nucleotide complexes wherein the nucleotide is associated with a mixture of divalent and monovalent cations. Preferably, the divalent cation may be magnesium or manganese.

The present invention relates to an improved process for synthesis of deoxyribonucleic acid (DNA), in particular cell-free enzymatic synthesis of DNA, preferably on a large or industrial scale, with an improved yield and/or with an improved efficiency. The invention requires the use of nucleotide complexes wherein the nucleotide is associated with a mixture of divalent and monovalent cations. Preferably, the divalent cation may be magnesium or manganese.

Embodiments of the present disclosure relate to method of preparing a substrate for sequencing by synthesis, including capturing library DNA to the surface using a low salt buffer solution prior to grafting primer oligonucleotides. Substrates prepared by the method described herein have increased monoclonality of clusters and sequencing by synthesis using the substrate prepared by the method are also described.

Embodiments of the present disclosure relate to method of preparing a substrate for sequencing by synthesis, including capturing library DNA to the surface using a low salt buffer solution prior to grafting primer oligonucleotides. Substrates prepared by the method described herein have increased monoclonality of clusters and sequencing by synthesis using the substrate prepared by the method are also described.

Disclosed herein is a more sensitive and accurate method of monitoring the pH of a solution, wherein the pH of the solution is quantified as a function of the electrochemical response of the solution in a two or three-electrode electrochemical cell, wherein the solution comprises a compound capable of undergoing a change in its oxidation state and/or structural conformation as a function of the pH of the solution. Also disclosed are highly accelerated methods and processes enabling analysis of specific polynucleotide sequences in a sample, e.g. a biological sample. The methods disclosed herein are, for example, useful for rapid screening of a large amount of samples in a point-of-care setting.

Disclosed herein is a more sensitive and accurate method of monitoring the pH of a solution, wherein the pH of the solution is quantified as a function of the electrochemical response of the solution in a two or three-electrode electrochemical cell, wherein the solution comprises a compound capable of undergoing a change in its oxidation state and/or structural conformation as a function of the pH of the solution. Also disclosed are highly accelerated methods and processes enabling analysis of specific polynucleotide sequences in a sample, e.g. a biological sample. The methods disclosed herein are, for example, useful for rapid screening of a large amount of samples in a point-of-care setting.

Compositions and methods are disclosed for substantially liquid, gel, suspension, slurry, semisolid and/or colloid storage of biological samples following admixture with the herein disclosed storage composition, permitting substantial recovery of biological activity following storage without refrigeration. In certain embodiments, unfractionated saliva samples may be stored without refrigeration for weeks, months or years in a form that permits recovery of intact DNA following the storage period.

Compositions and methods are disclosed for substantially liquid, gel, suspension, slurry, semisolid and/or colloid storage of biological samples following admixture with the herein disclosed storage composition, permitting substantial recovery of biological activity following storage without refrigeration. In certain embodiments, unfractionated saliva samples may be stored without refrigeration for weeks, months or years in a form that permits recovery of intact DNA following the storage period.