Invention Grant
- Patent Title: Method and kit for detecting a wild-type and/or a mutated target DNA sequence
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Application No.: US14439859Application Date: 2013-10-31
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Publication No.: US09938574B2Publication Date: 2018-04-10
- Inventor: Francesca Fontana , Nicolo Manaresi
- Applicant: Menarini Silicon Biosystems S.p.A.
- Applicant Address: IT Bologna
- Assignee: Menarini Silicon Biosystems, S.p.A.
- Current Assignee: Menarini Silicon Biosystems, S.p.A.
- Current Assignee Address: IT Bologna
- Agency: Dorsey & Whitney LLP
- Priority: ITTO2012A0962 20121031
- International Application: PCT/IB2013/059827 WO 20131031
- International Announcement: WO2014/068519 WO 20140508
- Main IPC: C12Q1/68
- IPC: C12Q1/68

Abstract:
The present invention relates to a method for detecting a first and/or a second target DNA sequence from a DNA library, differing in that a mutation generates/eliminates a restriction site for a restriction endonuclease, comprising the steps of: (a) providing the DNA library, in which each of the DNA sequences comprises a first sequence segment, a second sequence segment of genomic DNA as cleaved by the restriction endonuclease, and a third sequence segment reverse complementary to the union of said first sequence segment and 5′ overhang, if any, of the restriction endonuclease; (b) amplifying the library of DNA sequences by PCR using: a first reverse primer which hybridizes to the 3′ end region of the second sequence segment of the first or second target sequence positive strand; a second forward primer which hybridizes to the 3′ end region of the second sequence segment of the first target sequence antipositive strand; a third forward primer comprising a first portion hybridizing to the 5′ end region of the third sequence segment of the second target sequence antipositive strand and a second portion hybridizing to the 3′ end region of the second sequence segment of the second target sequence antipositive strand, wherein the first portion of the third forward primer has a length from 20% to 80% with respect to the total length of the third forward primer; (c) detecting DNA sequences amplified in step (b).
Public/Granted literature
- US20150368709A1 Method and Kit for Detecting a Wild-Type and/or a Mutated Target DNA Sequence Public/Granted day:2015-12-24
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