Patent classifications
C12N15/1068
METHOD FOR CONSTRUCTING SINGLE CELL SEQUENCING LIBRARY AND USE THEREOF
Provided in the present invention is a method for constructing a single cell sequencing library, comprising the following steps: a) lysing a single cell to obtain a single cell lysate; b) separating the nucleus and the cytoplasm in the single cell lysate obtained in step a) to obtain a nuclear solution and a total RNA solution; and c) constructing a chromatin DNA library with the nuclear solution obtained in step b) to obtain a chromatin-accessibility sequencing library of the single cell; and constructing a transcriptome library with the total RNA solution obtained in step b) to obtain a transcriptome sequencing library of the single cell.
Optical methods and systems for DNA assembly for computer data storage
An array-based system of assembled DNA for computer data storage is described. An array surface contains immobilized seed DNA initially having with blunt (or blocked) ends with a photocleavable optical linker at a forward end thereof holding the last few base pairs. A light source is light is applied to break the linker, generating a sticky end which allows for hybridization. Data-bearing DNA cassettes are introduced to the array and attach via their sticky ends to the unblock sites on the array surface. The attachment is made permanent via ligase.
Method for obtaining structural information concerning an encoded molecule and method for selecting compounds
In one aspect, the present invention relates to a method for obtaining structural information about an encoded molecule. The encoded molecule may be produced by a reaction of a plurality of chemical entities and may be capable of being connected to an identifier oligonucleotide containing codons informative of the identity of the chemical entities which have participated in the formation of the encoded molecule. In a certain embodiment, primers are designed complementary to the codons appearing on the identifier oligonucleotide, and the presence, absence or relative abundance of a codon is evaluated by mixing a primer with the identifier oligonucleotide in the presence of a polymerase and substrate (deoxy)ribonucleotide triphosphates measuring the extension reaction. In another aspect, the invention provides a method for selecting compounds which binds to a target. More specifically, the invention relates to a method in which a target associated with an oligonucleotide initially is mixed with a library of complexes, each complex comprising a display molecule and an oligonucleotide identifying said display molecule. Next, due an increased proximity, the target oligonucleotide is coupled to the identifier oligonucleotide of complexes having a display molecule with affinity towards the target. In a final stage the coupled nucleotides are analyzed to deduce at least the identity of the display molecule.
Methods and reagents for enhanced next generation sequencing library conversion and incorporation of molecular barcodes into targeted and random nucleic acid sequences
Novel engineered compositions, reagents, and methods are described that facilitate NGS analysis of both random and specific nucleic acid sequences in a sample by providing high efficiency target enrichment and improved error suppression. Specifically, the design and use of engineered NGS dual adapter molecules with homology regions (HRs) targeted at the 5 and 3 regions of a double-stranded DNA target, unique molecule identifiers (UMIs), and NGS adapters allows target libraries to be created for subsequent NGS analysis.
ENZYMATIC ENCODING METHODS FOR EFFICIENT SYNTHESIS OF LARGE LIBRARIES
- Thomas Franch ,
- Mikkel Dybro LUNDORF ,
- Søren Nyboe Jakobsen ,
- Eva Kampmann Olsen ,
- Anne Lee Andersen ,
- Anette Holtmann ,
- Anders Holm HANSEN ,
- Anders Malling SØRENSEN ,
- Anne GOLDBECH ,
- Daen de LEON ,
- Ditte Kievsmose KALDOR ,
- Frank Abildgaard Sløk ,
- Gitte Nystrup Husemoen ,
- Johannes DOLBERG ,
- Kim Birkebæk Jensen ,
- Lene PETERSEN ,
- Mads Nørregaard-Madsen ,
- Michael Anders Godskesen ,
- Sanne Schrøder Glad ,
- Søren Neve ,
- Thomas Thisted ,
- Tine Titilola Akinleminu KRONBORG ,
- Christian Klarner SAMS ,
- Jakob Felding ,
- Per-Ola Freskgard ,
- Alex GOULIAEV ,
- Henrik Pedersen
Disclosed is a method for obtaining a bifunctional complex comprising a molecule linked to a single stranded identifier oligonucleotide, wherein a nascent bifunctional complex comprising a chemical reaction site and a priming site for enzymatic addition of a tag is a) reacted at the chemical reaction site with one or more reactants, and b) reacted enzymatically at the priming site with one or more tag(s) identifying the reactant(s).
Method of constructing sequencing library
Provided is a method of constructing a sequencing library. The method includes 1) providing a single-stranded DNA fragment from a biological sample; 2) subjecting the single-stranded DNA fragment to whole genomic amplification to obtain a whole genome amplification product; 3) fragmenting the whole genome amplification product using a transposase embedded with two adaptors to obtain a fragmented product with two adaptors respectively at two ends; and 4) amplifying the fragmented product with two adaptors respectively at two ends using a tag sequence and a pair of primers to obtain said sequencing library.
ANALYSIS SYSTEM FOR ORTHOGONAL ACCESS TO AND TAGGING OF BIOMOLECULES IN CELLULAR COMPARTMENTS
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.
Microarray synthesis and assembly of gene-length polynucleotides
There is disclosed a process for in vitro synthesis and assembly of long, gene-length polynucleotides based upon assembly of multiple shorter oligonucleotides synthesized in situ on a microarray platform. Specifically, there is disclosed a process for in situ synthesis of oligonucleotide fragments on a solid phase microarray platform and subsequent, on device assembly of larger polynucleotides composed of a plurality of shorter oligonucleotide fragments.
Methods For Directed Folding Assembly Or Dimerization Of Proteins By Templated Assembly Reactions
The present disclosure provides nucleic acid molecules, compositions, and kits comprising the same, and methods for producing templated assembly products.
Method and system for sequencing nucleic acids
Provided are compositions, methods and systems for determining the sequence of a template nucleic acid using a polymerase-based, sequencing-by-binding procedure. An examination step involves monitoring the interaction between a polymerase and template nucleic acid in the presence of one or more nucleotides. Identity of the next correct nucleotide in the sequence is determined without incorporation of any nucleotide into the structure of the primer by formation of a phosphodiester bond. An optional incorporation step can be used after the examination step to extend the primer by one or more nucleotides, thereby incrementing the template nucleotides that can be examined in a subsequent examination step. The sequencing-by-binding procedure does not require the use of labeled nucleotides or polymerases, but optionally can employ these reagents.