Provided herein are compositions and methods for accurate and scalable single cell multiomics methods, and their applications for mutational analysis in research. diagnostics, and treatment. Further provided herein are multiomics methods for parallel analysis of DNA, RNA, and/or proteins from single cells using Primary Template-Directed Amplification (PTA) nucleic acid amplification.

A method for producing pancreatic endocrine cells, including introducing (B) or (D) below into cultured somatic cells to transdifferentiate into a population of pancreatic endocrine cells that include insulin-producing beta-cells: (B) a GLIS1 gene or one or more gene products thereof, a Neurogenin3 gene or one or more gene products thereof, and a Pdx1 gene or one or more gene products thereof; and (D) a GLIS1 gene or one or more gene products thereof, a Neurogenin3 gene or one or more gene products thereof, and a MafA gene or one or more gene products thereof. The GLIS1 gene includes the nucleotide sequence as set forth in SEQ ID NO: 1 or SEQ ID NO: 2. The population of pancreatic endocrine cells are produced without undergoing an iPS cell stage.

The invention is directed to methods for massively parallel template-free enzymatic synthesis of a plurality of different polynucleotides of predetermined sequences. In one aspect, methods of the invention employ large scale arrays of reaction sites each associated with at least one working electrode for controlling deprotection and deblocking steps at predetermined user selected sites. In another aspect, the invention provides template-free enzymatic synthesis with proofreading, wherein completed polynucleotides at predetermined reaction sites are sequenced using a sequencing by synthesis technique, particularly employing electrochemically labile blocking groups.

Provided are a method for purifying a nucleic acid library, and a kit, the method comprising the steps of: providing a nucleic acid library comprising single-stranded template nucleic acids; obtaining a library of complementary nucleic acids by binding complementary nucleic acid units to each base of the strand of the template nucleic acids; introducing at least one modified nucleic acid unit during the binding process of the nucleic acid units; and selectively selecting a nucleic acid having a desired length from the library of complementary nucleic acids using the modified nucleic acid unit. According to the present invention, the nucleic acid library may be purified regardless of the complexity, sequence or length of the nucleic acid library, and nucleic acids having different lengths may be simultaneously purified. The purification may be carried out through direct experiment or using a next-generation sequencing instrument.

The invention relates to a system and methods for enhancing access to nuclear informational molecules, such as DNA, RNA, and proteins, by analytical biomolecules, such as transposome complexes, by treating nuclei with a nuclear permeability enhancer, and to methods of using nuclear membrane, cell membrane, and external compartmentalization approaches as contiguity preserving elements.

The present disclosure relates to precursor molecules of DNA-encoded compounds, and methods of preparing thereof. In some aspects, provided herein are methods of synthesizing DNA-encoded compounds, and libraries thereof, from precursor molecules and positional building blocks.

Disclosed are methods for detecting a target molecule in a test sample using a cell-free protein synthesis (CFPS) reaction. The methods may be utilized for detecting target molecules which may include small molecules and/or metabolites of small molecules. The methods employ one or more transcription templates that encode and conditionally express one or more exogenous RNA polymerases in the presence of the target molecule. The expressed RNA polymerases in turn induce expression of one or more reporter molecules from transcription templates comprising promoters for the RNA polymerases, thereby amplifying an output signal that is generated in the presence of a detected target molecule.

A transposome complex capable of producing size-controlled nucleic acid fragments is described herein. In some embodiments, the transposome complex includes multiple inactive transposomes with active transposomes on both ends of the multiple inactive transposomes. Applications, uses, and variations of the disclosed transposome complex include, but are not limited to, library preparation for a nucleic acid and tuning the length of the transposome complex to produce nucleic acid fragments of predetermined or desired lengths.

Disclosed is a DNA assembly mix, comprising a 3-5 exonuclease enzyme which is XthA; and a buffer. Also disclosed is a DNA assembly mix, comprising a polymerase and ligase free composition comprising a 3-5 exonuclease enzyme; and a buffer. Also disclosed is a method of assembling a plurality of DNA fragments, comprising: (a) mixing the plurality of DNA fragments with the DNA assembly mix as disclosed herein; and (b) incubating the mixture from step (a) at a temperature for a period of time suitable for assembling the plurality of DNA fragments. Further disclosed is use of the DNA assembly mix as disclosed herein in high-throughput DNA assembly, wherein the DNA assembly mix is used in a microfluidic platform to assemble DNA.

The present disclosure relates to bivalent or polyvalent linear initiator nucleic acids comprising initial building blocks and a coding region. The linear initiator nucleic acids may be used for the synthesis of an encoded compound to produce bivalent or polyvalent molecules.