EXPONENTIAL BASE-3 AND GREATER NUCLEIC ACID AMPLIFICATION WITH REDUCED AMPLIFICATION TIME

    公开(公告)号:US20200239878A1

    公开(公告)日:2020-07-30

    申请号:US16667446

    申请日:2019-10-29

    申请人: Cepheid

    发明人: Russell Higuchi

    摘要: Described herein are methods and compositions that provide highly efficient nucleic acid amplification. In some embodiments, this allows a 3-fold or greater increase of amplification product for each amplification cycle and therefore increased sensitivity and speed over conventional PCR. Modified bases can be employed in primers to provide this base-3 or greater amplification with satisfactory PCR cycle times, which are improved, as compared to those observed in the absence of modified bases.

    Integrated purification and measurement of DNA methylation and co-measurement of mutations and/or mRNA expression levels in an automated reaction cartridge

    公开(公告)号:US11260387B2

    公开(公告)日:2022-03-01

    申请号:US15839731

    申请日:2017-12-12

    申请人: Cepheid

    摘要: 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) desulphonating the bound deaminated nucleic acid and/or simultaneously eluting and desulphonating 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.

    EXPONENTIAL BASE-GREATER-THAN-2 NUCLEIC ACID AMPLIFICATION

    公开(公告)号:US20210395797A1

    公开(公告)日:2021-12-23

    申请号:US17306761

    申请日:2021-05-03

    申请人: Cepheid

    发明人: Russell Higuchi

    IPC分类号: C12Q1/686

    摘要: Described herein are methods and compositions that provide highly efficient nucleic acid amplification. In some embodiments, this allows a greater than 2-fold increase of amplification product for each amplification cycle and therefore increased sensitivity and speed over conventional PCR.

    Exponential base-greater-than-2 nucleic acid amplification

    公开(公告)号:US11028434B2

    公开(公告)日:2021-06-08

    申请号:US16359947

    申请日:2019-03-20

    申请人: Cepheid

    发明人: Russell Higuchi

    IPC分类号: C12Q1/68 C12Q1/686

    摘要: Described herein are methods and compositions that provide highly efficient nucleic acid amplification. In some embodiments, this allows a greater than 2-fold increase of amplification product for each amplification cycle and therefore increased sensitivity and speed over conventional PCR.