Patent classifications
C12N15/1006
Desorption of nucleic acids
In a method for the desorption of nucleic acids from a sample, in order to simplify the desorption of nucleic acids from the sample, a solid phase is repeatedly rinsed with an elution buffer in a microfluidic system, in order to elute nucleic acids bonded to the solid phase from the solid phase in the microfluidic system.
ENHANCED PURIFICATION OF ADENO-ASSOCIATED VIRUS TO MORE EFFECTIVELY REMOVE CONTAMINATING DNA
A method for reducing a contaminating DNA content of a preparation containing AAV capsids and contaminating DNA, comprising the steps of a) Performing an extraction of DNA with a solid phase bearing positive charges at its surface said solid phase is contacted with the preparation at a pH of 7.0±1.0, and a salt concentration of 10 mM to 200 mM yielding a first fraction, (b) Diafiltering the first fraction by a first tangential flow filtration to obtain a second fraction, (c) Treating the second fraction with DNase, (d) Diafiltering the DNase treated second fraction obtained by step c) by a second tangential flow, (e) filtration to a buffer with pH of 7.0±1.0, and a salt concentration of 10 mM to 20 mM to yield a third fraction, and optionally (f) Concentrating the third fraction by tangential flow filtration before supplemental chromatography.
METHOD FOR IDENTIFYING RNA BINDING PROTEIN BINDING SITES ON RNA
The invention relates to methods for purifying and isolating at least one RNA molecule which interacts with an RNA-binding protein (RBP). The invention also provides nucleic acid adaptors and primers for use in such methods.
Process for concentrating cells from a sample and then isolating nucleic acids from said cells
Biological cells are concentrated and isolated from a sample and/or biological cells are concentrated from a sample, followed by the isolation of nucleic acids from said cells. The sample is brought into contact with a solid phase which has a rough or structured surface.
Single cell cellular component enrichment from barcoded sequencing libraries
The present invention relates to the analysis of complex single cell sequencing libraries. Disclosed are methods for enrichment of library members based on the presence of cell-of origin barcodes to identify and concentrate DNA that is relevant to interesting cells or components that would be expensive or difficult to study otherwise. Also, disclosed are methods of capturing cDNA library molecules by use of CRISPR systems, hybridization or PCR. The present invention allows for identifying the properties of rare cells in single cell RNA-seq data and accurately profile them through clustering approaches. Further information on transcript abundances from subpopulations of single cells can be analyzed at a lower sequencing effort. The methods also allow for linking TCR alpha and beta chains at the single cell level.
Single cell cellular component enrichment from barcoded sequencing libraries
The present invention relates to the analysis of complex single cell sequencing libraries. Disclosed are methods for enrichment of library members based on the presence of cell-of origin barcodes to identify and concentrate DNA that is relevant to interesting cells or components that would be expensive or difficult to study otherwise. Also, disclosed are methods of capturing cDNA library molecules by use of CRISPR systems, hybridization or PCR. The present invention allows for identifying the properties of rare cells in single cell RNA-seq data and accurately profile them through clustering approaches. Further information on transcript abundances from subpopulations of single cells can be analyzed at a lower sequencing effort. The methods also allow for linking TCR alpha and beta chains at the single cell level.
HIGH THROUGHPUT LINKING OF MULTIPLE TRANSCRIPTS
Provided are high throughput methods for physically linking cDNA molecules derived from mRNA molecules expressed by the same cell, and libraries of linked cDNA molecules produced by the methods. The methods comprise reverse transcribing mRNA from a single cell in a first container to produce cDNA molecules, and linking the cDNA molecules in a second container. The methods unexpectedly produced libraries of cDNA molecules with an increase in the number of molecules that are correctly linked to other molecules derived from the same cell.
AFFINITY PURIFICATION SEQUENCING
Described herein are affinity-labeled polypeptide compositions, such as affinity-labeled transcription factor compositions, and methods of using such compositions to evaluate interactions of the polypeptide with other molecules such as nucleic acids.
METHOD AND MATERIALS FOR ISOLATION OF NUCLEIC ACID MATERIALS
A method for nucleic acid isolation comprising: receiving a binding moiety solution within a process chamber; mixing the binding moiety solution with a biological sample, within the process chamber, in order to produce a moiety-sample mixture; incubating the moiety-sample mixture during a time window, thereby producing a solution comprising a set of moiety-bound nucleic acid particles and a waste volume; separating the set of moiety-bound nucleic acid particles from the waste volume; washing the set of moiety-bound nucleic acid particles; and releasing a nucleic acid sample from the set of moiety-bound nucleic acid particles. The method preferably utilizes a binding moiety comprising at least one of poly(allylamine) and polypropylenimine tetramine dendrimer, both of which reversibly bind and unbind to nucleic acids based upon environmental pH.
Method for isolating a target nucleic acid including small target nucleic acids with high yield
The present invention pertains to a method for isolating a target nucleic acid including small target nucleic acids from a sample, said method comprising at least the following steps a) binding at least a portion of the target nucleic acid including small target nucleic acids to a nucleic acid binding solid phase comprised in a column by passing the sample through said column, b) performing an enzymatic and/or chemical treatment on the nucleic acid binding solid phase while the target nucleic acid is bound to said solid phase, c) collecting at least a portion of the small target nucleic acids released from the solid phase during said treatment of step b) as flow-through, d) contacting said flow-through which comprises small target nucleic acids mixed with a recovery solution with a nucleic acid binding solid phase for binding the contained small target nucleic acids to said nucleic acid binding solid phase, e) optionally performing an elution. The present invention results in a considerable increase in the yield of small target nucleic acids in the isolated target nucleic acid because it allows to efficiently capture and recover small target nucleic acids.