公开(公告)号：US11603555B2

公开(公告)日：2023-03-14

申请号：US15182394

申请日：2016-06-14

Applicant: Cepheid

Inventor： Edwin W. Lai ,  Andrew Kohlway ,  Reuel Vanatta ,  Russell Higuchi ,  Alexander Gall ,  Kriszten Kocmond

IPC: C12Q1/6806 ,  C12Q1/6827 ,  C12Q1/6886 ,  C12N15/10 ,  B01L7/00

Abstract: In various embodiments methods of determining methylation of DNA are provided. In one illustrative, but non-limiting embodiment the method comprises i) contacting a biological sample comprising a nucleic acid to a first matrix material comprising a first column or filter where said matrix material binds and/or filters nucleic acids in said sample and thereby purifies the DNA; ii) eluting the bound DNA from the first matrix material and denaturing the DNA to produce eluted denatured DNA; iii) heating the eluted DNA in the presence of bisulfite ions to produce a deaminated nucleic acid; iv) contacting said deaminated nucleic acid to a second matrix material comprising a second column to bind said deaminated nucleic acid to said second matrix material; v) desulfonating the bound deaminated nucleic acid and/or simultaneously eluting and desulfonating the nucleic acid by contacting the deaminated nucleic acid with an alkaline solution to produce a bisulfite converted nucleic acid; vi) eluting said bisulfite converted nucleic acid from said second matrix material; and vii) performing methylation specific PCR and/or nucleic acid sequencing, and/or high resolution melting analysis (HRM) on said bisulfite-converted nucleic acid to determine the methylation of said nucleic acid, wherein at least steps iv) through vi) are performed in a single reaction cartridge.

公开(公告)号：US20150252354A1

公开(公告)日：2015-09-10

申请号：US14431243

申请日：2013-09-26

Applicant: CEPHEID

Inventor： Edwin Wei-Lung Lai ,  Reuel Vanatta ,  Kenneth E. Ho

IPC: C12N15/10 ,  G06F19/20 ,  C12Q1/68

CPC classification number: C12N15/1003 ,  C12Q1/6806 ,  C12Q1/6886 ,  C12Q2600/118 ,  C12Q2600/158 ,  C12Q2600/16 ,  G01N1/06 ,  G06F19/20

Abstract: Methods and reagents are provided for the rapid extraction of nucleic acids from a fixed paraffin embedded sample (e.g., a FFPET sample). In some embodiments, the methods comprise incubating one or more sections of said tissue sample in a lysis solution comprising a buffer sufficient to maintain the pH of said solution at a pH ranging from about pH 4 to about pH 9; a chaotropic agent; a chelating agent; and a detergent; where the incubating is at a temperature ranging from about 50 C to about 100 C; and recovering the nucleic acid from said lysis solution.

Abstract translation: 提供方法和试剂用于从固定石蜡包埋样品（例如，FFPET样品）中快速提取核酸。 在一些实施方案中，所述方法包括将所述组织样品的一个或多个切片孵育在包含足以将所述溶液的pH保持在约pH 4至约pH 9范围内的pH的溶解溶液中; 离液剂; 螯合剂; 和洗涤剂; 其中孵育温度为约50℃至约100℃; 并从所述裂解液中回收核酸。