Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Statistical analysis for noninvasive chromosomal aneuploidy determination

An aneuploidy, X-chromosome technique used in the field of statistical analysis for non-invasive sex chromosome aneuploidy determination

Active Publication Date: 2020-10-02
F HOFFMANN LA ROCHE & CO AG
View PDF108 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention addresses this need

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
  • Statistical analysis for noninvasive chromosomal aneuploidy determination
  • Statistical analysis for noninvasive chromosomal aneuploidy determination
  • Statistical analysis for noninvasive chromosomal aneuploidy determination

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0232] Example 1: Preparation of DNA for the Tandem Ligation Procedure

[0233] Genomic DNA from subjects was obtained from Coriell Cell Repositories (Camden, New Jersey) and fragmented by acoustic shearing (Covaris, Woburn, MA) to an average fragment size of approximately 200 bp.

[0234] DNA was biotinylated using standard procedures. Briefly, Covaris fragmented DNA ends were repaired by generating the following reactions in 1.5 ml microtubes. 5ug DNA, 12 μl 10x T4 ligase buffer (Enzymatics, Beverly MA), 50 U T4 polynucleotide kinase (Enzymatics, Beverly MA) and H 2 0 to 120 μl. The microtubes were incubated at 37°C for 30 minutes. DNA was diluted with 10 mM Tris 1 mM EDTA pH 8.5 to a desired final concentration of ~0.5 ng / μl.

[0235] 5 μl of DNA was placed in each well of a 96-well plate, and the plate was sealed with adhesive parafilm and spun at 250×g for 10 seconds. The plate was then incubated at 95°C for 3 minutes, cooled to 25°C, and spun again at 250 xg for 10 ...

Embodiment 2

[0241] Example 2: Universal amplification of ligated products

[0242] The polymerized and / ligated nucleic acid is amplified using universal PCR primers complementary to the universal sequence present in the first and second fixed sequence oligonucleotides that hybridize to the nucleic acid region of interest. 25 μl of each of the reaction mixtures in Example 3 were used in each amplification reaction. 50 μL Universal PCR consisting of 25 μL of eluted ligation product plus 1x Pfusion buffer (Finnzymes, Finland), 1 M betaine, 400 nM of each dNTP, 1 U Pfusion error-correcting thermostable DNA polymerase, and primer pairs with sample tags Reactions are used to uniquely identify individual samples prior to pooling and sequencing. Using BioRad Tetrad TM Thermal cyclers perform PCR under stringent conditions.

[0243] 10 μl of universal PCR products from each of the samples were pooled and quantified using Quant-iT TM PicoGreen, (Invitrogen, Carlsbad, CA) purified and quantifie...

Embodiment 3

[0244] Example 3: Analysis of polymorphic loci to assess percent fetal contribution

[0245] To assess the proportion of fetal nucleic acid in a maternal sample, an assay was designed for a panel of SNP-containing loci on chromosomes 1 to 12, in which two bridging oligonucleotides differing by one base were used to interrogate each SNP (see, e.g. image 3 ). SNPs were optimized for minor allele frequency in the HapMap 3 dataset. Duan, et al., Bioinformation, 3(3):139-41 (2008); Epub 2008 Nov 9.

[0246] Oligonucleotides were synthesized by IDT (Coralville, Iowa) and pooled together to generate a single multiplexed assay pool. PCR products were generated from each subject sample as previously described. An informative polymorphic locus is defined as a locus in which the fetal allele differs from the maternal allele. Because the assay exhibits an allele specificity of over 99%, informative loci are readily identified when the fetal allelic ratio of the locus is measured to b...

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 provides non-invasive determination of X and / or Y chromosome abnormalities indicative of aneuploidy or gender mosaicism in a maternal sample by detecting and determining the relative contribution of genetic sequences from the X chromosome and / or Y chromosome in a maternal sample Methods.

Description

[0001] priority [0002] This application claims the benefit of U.S. Patent Application Serial No. 13 / 917,329, filed June 13, 2013, which is based on U.S. Patent Application Serial No. 13 / 790,642, filed March 8, 2013 Continuation-in-Part and is a continuation-in-part of U.S. Patent Application Serial No. 13 / 338,963 filed December 28, 2011, which is a U.S. Patent Application Serial No. 13 / 338,963 filed December 9, 2011 Continuation-in-Part Application Serial No. 13 / 316,154, U.S. Patent Application Serial No. 13 / 316,154 claims priority to U.S. Provisional Patent Application Serial No. 61 / 436,135, filed January 25, 2011, each of which is assigned to the present invention the assignee. [0003] field of invention [0004] The present invention relates to non-invasive sex determination of fetal or statistical analysis of X and Y chromosome frequency abnormalities by detecting and determining the relative contribution of genetic sequences from the X and Y chromosomes in view of the ...

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 Patents(China)
IPC IPC(8): C12Q1/6883G16B20/10G16B20/20
CPCC12Q1/6883C12Q2600/156G16B20/00G16B20/20G16B20/10
Inventor 克雷格·斯特鲁布尔阿诺德·奥利芬特埃里克·王
Owner F HOFFMANN LA ROCHE & CO AG
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com