Siemens Healthcare Gmbh

Siemens Aktiengesellschaft

Siemens Healthineers Ag

Siemens Healthcare Diagnostics Gmbh

Yiwei Huang

Walter Gumbrecht

Peter Paulicka

Ramya Vutukuru

Divya Khandige Sharma

Ragavendar Ms

Nivedita Mitra

Carsten Dietrich

Mark Matzas

Maximilian Würstle

Andreas Emanuel Posch

Tivadar Mach

Stefan Prause

DIRECT RNA NANOPORE SEQUENCING WITH HELP OF A STEM-LOOP REVERSE POLYNUCLEOTIDE

The present invention relates to a method of selectively amplifying at least one nucleic acid sequence of at least one microorganism and/or virus in a sample of a subject, wherein k-mers 3 are applied that show a difference in frequency and/or context in the genome 2 of the at least one microorganism and/or virus compared to the genome of the subject 1.

Amplification-integrated genetic material depletion of non-target organisms using differentially abundant k-mers

The present invention relates to a method of obtaining an enriched population of a target polynucleotide using a synthetic single guide RNA (sgRNA) for an sgRNA-guided nucleic acid-binding protein, as well as to a method of obtaining a pool of target-irrelevant synthetic single guide RNAs (sgRNAs) for a sgRNA-guided nucleic acid-binding protein. Also provided is a target polynucleotide and sgRNAs obtainable by the methods of the invention. Further envisaged is a kit comprising a pool of sgRNAs obtainable by the method of the invention, and the use of a pool of sgRNAs obtainable by the methods of the invention.

Target irrelevant guide RNA for CRISPR

A method includes (i) providing an RNA polynucleotide; (ii) modifying the RNA polynucleotide by annealing and ligating a polynucleotide comprising a 3' terminal random multimer segment and having a stem-loop form; (iii) optionally performing a reverse transcription of the RNA polynucleotide; (iv) cleaving the stem-loop segment of the annealed polynucleotide to yield a 3&#x2032; A overhang; (v) connecting an adaptor polynucleotide complex associated with an RNA translocase enzyme and at least one cholesterol tether segment to the polynucleotide obtained in step (iv); (vi) contacting the modified RNA polynucleotide obtained in step (v) with a transmembrane pore such that the RNA translocase controls the movement of the RNA polynucleotide through the transmembrane pore and the cholesterol tether anchors the RNA polynucleotide in the vicinity of the transmembrane pore; and (vii) taking one or more measurements during the movement of the RNA polynucleotide through the transmembrane pore Other features are also disclosed.

Direct RNA nanopore sequencing with help of a stem-loop reverse polynucleotide

A method and apparatus for enrichment and detection of low abundance pathogens are provided. The method includes adding one or more proteins containing pathogen binding domains to the sample. The sample is incubated to form a complex thereby. The complex is separated from the sample, and an apparatus is provided for enriching pathogen DNA that achieves pathogen detection levels as low as 1 cfu/ml. The method further includes adding a lysis buffer to the separated complex and incubating to form a mixture. A buffer containing guanidine thiocyanate is added to the mixture. The mixture is connected with a matrix to form a bound entity, and the bound entity is separated from the mixture.

Method and apparatus for enriching pathogen DNA

A method and an apparatus are provided for enriching pathogen DNA. The method includes adding a selective lysis buffer to a sample, incubating the mixture formed thereby, and filtering the mixture. The apparatus for enriching pathogen DNA includes: a lysis chamber; a reservoir containing selective lysis buffer, connected to the lysis chamber; and a filter connected to the lysis chamber; that achieves a limit of detection of pathogens of 500 cfu/ml or less.

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Erlangen, Germany

Yiwei Huang