Unlock instant, AI-driven research and patent intelligence for your innovation.

Monotube preparation of DNA and RNA for sequencing

A sequencing and target technology, applied in the field of nucleic acid sequencing and enrichment of rare nucleic acid targets for sequencing, can solve the problems of waste of sample materials, labor and reagent costs, etc.

Pending Publication Date: 2022-04-01
F HOFFMANN LA ROCHE & CO AG
View PDF11 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This results in wasted sample material as well as labor and reagent costs

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Monotube preparation of DNA and RNA for sequencing
  • Monotube preparation of DNA and RNA for sequencing
  • Monotube preparation of DNA and RNA for sequencing

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0058] Example 1. Simultaneous preparation of sequence-ready DNA and RNA from cell line samples

[0059] to generate Figure 4Based on the data described in , an optimized DNA / RNA workflow was used for the mixing of cell line DNA / cell line RNA with cell lines containing known fusions (EML4-ALK and SLC34A2-ROS1 fusions). Gene-specific primers were used to target relevant exons in ALK and ROS1 during reverse transcription. As shown above, reverse transcription proceeds in such a way that the DNA retains its library-ready capacity (ie remains duplex and relatively intact). Following reverse transcription, RNAseH treatment, samples were taken into the end repair / A-tailing step of the AVENIO Tumor Tissue Analysis Kit workflow (Roche Sequencing Solutions, Inc. Pleasanton, CA) and AVENIO was used for the remainder of the workflow. Tumor tissue protocol execution. The goal of this experiment was to determine whether RNA fusions could be detected while maintaining the expected depth...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The present invention is methods and compositions for simultaneously forming libraries for nucleic acid sequencing from DNA and RNA present in a sample.

Description

technical field [0001] The invention relates to the field of nucleic acid sequencing. More specifically, the invention relates to the field of enrichment of rare nucleic acid targets for sequencing. Background technique [0002] Many workflows exist for the separate preparation and sequencing of DNA and RNA samples. When information needs to be obtained from both DNA and RNA, some workflows are completely independent, while others may be joined as one workflow at some step in the process. In either case, if information is required from both DNA and RNA from a single source (such as a patient sample), two separate samples are required, one for isolating DNA and another for isolating RNA. This results in waste of sample material as well as labor and reagent costs. For precious samples, such as clinical biopsy samples, forensic samples, or historical samples, there may not be enough material to perform two separate isolations of DNA and RNA. Combined DNA and RNA workflows w...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12Q1/6806
CPCC12Q1/6806C12N15/1072C12N15/1096C12N15/1093C12Q2521/107C12Q2525/161C12Q2525/191C12Q2535/122C12Q2563/179C12Q1/6855C12Q1/6874
Inventor D·M·克拉斯A·洛夫乔伊M·洛伊泽K·M·麦克林托克
Owner F HOFFMANN LA ROCHE & CO AG