A surface functionalizing method for use in high-throughput in situ synthesis of nucleic acids by 3D inkjet printing. The method includes subjecting a surface of a substrate to hydroxyl enrichment treatment; adding hydrophobic molecules to the surface of the substrate, the hydrophobic molecules being not reactive with phosphoramidite monomers; spraying, by a multi-channel piezoelectric inkjet head assembly, an etching ink to a predetermined area on the surface of the substrate for micro-etching, the etching ink being prepared with a fluoride compound reactive with the hydrophobic molecules; and adding hydrophilic molecules to the surface of the substrate. By using the method, a functionalized surface with given areas being patterned can be formed on the surface of the substrate, and then a same multi-channel piezoelectric inkjet head assembly can be directly used for subsequent high-resolution printing of phosphoramidite monomers and synthesis of nucleic acids.

A surface functionalizing method for use in high-throughput in situ synthesis of nucleic acids by 3D inkjet printing. The method includes subjecting a surface of a substrate to hydroxyl enrichment treatment; adding hydrophobic molecules to the surface of the substrate, the hydrophobic molecules being not reactive with phosphoramidite monomers; spraying, by a multi-channel piezoelectric inkjet head assembly, an etching ink to a predetermined area on the surface of the substrate for micro-etching, the etching ink being prepared with a fluoride compound reactive with the hydrophobic molecules; and adding hydrophilic molecules to the surface of the substrate. By using the method, a functionalized surface with given areas being patterned can be formed on the surface of the substrate, and then a same multi-channel piezoelectric inkjet head assembly can be directly used for subsequent high-resolution printing of phosphoramidite monomers and synthesis of nucleic acids.

This invention discloses multi-part primers for primer-dependent nucleic acid amplification methods. Also disclosed are multiplex assay methods, related reagent kits, and oligonucleotides for such methods.

This invention discloses multi-part primers for primer-dependent nucleic acid amplification methods. Also disclosed are multiplex assay methods, related reagent kits, and oligonucleotides for such methods.

The present invention relates to an enzyme-catalyzed process for producing UDP-galactose from low-cost substrates uridine monophosphate and D-galactose in a single reaction mixture. Said process can be operated (semi)continuously or in batch mode. Said process can be extended to uridine as starting material instead of uridine monophosphate. Further, said process can be adapted to produce galactosylated molecules and biomolecules including saccharides, proteins, peptides, glycoproteins or glycopeptides, particularly human milk oligosaccharides (HMO) and (monoclonal) antibodies.

The present invention relates to an enzyme-catalyzed process for producing UDP-N-acetyl-α-D-glucosamine (UDP-GlcNAc) from low-cost substrates uridine monophosphate and N-acetyl-D glucosamine in a single reaction mixture with immobilized or preferably co-immobilized enzymes. Uridine may be used as starting material instead of uridine monophosphate as well. Further, said process may be adapted to produce GlcNAcylated molecules and biomolecules including saccharides, particularly human milk oligosaccharides (HMO), proteins, peptides, glycoproteins, particularly antibodies, or glycopeptides, and bioconjugates, particularly carbohydrate conjugate vaccines and antibody-drug conjugates.

The invention relates to a method for preparing nicotinamide mononucleotide by using nicotinamide as a raw material, which comprises: in acetonitrile, dichloromethane, 1,2-dichloroethane or liquid sulfur dioxide as a solvent, allowing nicotinamide and tetraacetyl ribose to react as catalyzed by trimethylsilyl trifluoromethanesulfonate or tin tetrachloride, adjusting a pH value thereof to 3-5, adding a sodium methoxide solution thereto to react at −15° C. to 5° C., adjusting a pH value thereof to 3-5, and subjecting the reaction mixture to microfiltration and nanofiltration using a membrane concentrator, thereby obtain a nicotinamide ribose solution; allowing the nicotinamide ribose solution to react as catalyzed by nicotinamide ribokinase in the presence of Mg ions, ATP and a buffer, thereby obtaining nicotinamide mononucleotide. The method of the invention omits the step of refining nicotinamide ribose, and thus has simpler process, lower cost and less time consumption, and has the advantages of faster reaction speed and lower enzyme consumption compared with the enzyme catalytic process directly using refined nicotinamide ribose solid.

Digestible primers are incorporated into single cell analysis workflows to reduce and/or eliminate primer byproducts and misprimed nucleic acids. Specifically, digestible primers can participate in a first reaction, such as reverse transcription of RNA transcripts to generate cDNA, but digestible primers are digested to prevent them from participating in subsequent reactions, such as nucleic acid amplification. For example, digestible primers can include a primer with one or more ribonucleotide nucleobases, a primer with uracil bases, a primer with deoxyuridine sequences, or a primer with ribouridine sequences. Such primers can then be digested (e.g., enzymatically digested) to remove them from interfering in subsequent nucleic acid amplification reactions.

The purpose of the present invention is to provide microorganisms which efficiently produce nicotinamide riboside, and microorganisms which can efficiently produce both nicotinamide mononucleotide and nicotinamide riboside. Nicotinamide mononucleotide and nicotinamide riboside can be produced by culturing lactic acid bacteria belonging to the genus Fructobacillus.

The purpose of the present invention is to provide microorganisms which efficiently produce nicotinamide riboside, and microorganisms which can efficiently produce both nicotinamide mononucleotide and nicotinamide riboside. Nicotinamide mononucleotide and nicotinamide riboside can be produced by culturing lactic acid bacteria belonging to the genus Fructobacillus.