Preparation of templates for methylation analysis
US-10954554-B2 · Mar 23, 2021 · US
Preparation of templates for methylation analysis
US12385085B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-12385085-B2 |
| Application number | US-202318500680-A |
| Country | US |
| Kind code | B2 |
| Filing date | Nov 2, 2023 |
| Priority date | Feb 7, 2007 |
| Publication date | Aug 12, 2025 |
| Grant date | Aug 12, 2025 |
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Abstract
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The invention relates to a method of preparing and using a library of template polynucleotides suitable for use as templates in solid-phase nucleic acid amplification and sequencing reactions to determine the methylation status of the cytosine bases in the library. In particular, the invention relates to a method of preparing and analysing a library of template polynucleotides suitable for methylation analysis.
First claim
Opening claim text (preview).
The invention claimed is: 1. A polynucleotide composition, comprising: adaptor-ligated double stranded nucleic acid target fragments, an individual adaptor-ligated double stranded nucleic acid target fragment of the adaptor-ligated double stranded nucleic acid target fragments comprising: a first adaptor polynucleotide and a second adaptor polynucleotide, wherein the first adaptor polynucleotide and the second adaptor polynucleotide each comprise a plurality of consecutive nucleotides, wherein all cytosine bases in the first adaptor polynucleotide and the second adaptor polynucleotide are methylated, and wherein first adaptor polynucleotide and the second adaptor polynucleotide each comprise a region of double stranded nucleic acids and at least one region of single stranded nucleic acids, the region of single-stranded nucleic acids forming a forked end, wherein the at least one region of single stranded nucleic acids comprises the plurality of consecutive nucleotides and wherein the plurality of consecutive nucleotides is complementary to the last 21 consecutive nucleotides of SEQ ID NO: 6; and a double-stranded target polynucleotide disposed between the first adaptor polynucleotide and the second adaptor polynucleotide. 2. The polynucleotide composition of claim 1 , wherein the double-stranded target polynucleotide is derived from genomic DNA. 3. The polynucleotide composition of claim 1 , wherein the double-stranded target polynucleotide comprises one or more methylated cytosines. 4. The polynucleotide composition of claim 1 , wherein the double-stranded target polynucleotide comprises one or more uracils. 5. The polynucleotide composition of claim 1 , comprising an amplification primer hybridized to the at least one region of single stranded nucleic acids. 6. The polynucleotide composition of claim 5 , wherein the amplification primer comprises a region that extends beyond a 5′ end of the at least one region of single stranded nucleic acids. 7. The polynucleotide composition of claim 1 , wherein one or both of the first adaptor polynucleotide or the second adaptor polynucleotide comprises a biotin group.
Assignees
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Classifications
for detection of mutation or polymorphism · CPC title
Methods for determination or identification of nucleic acids involving differential detection · CPC title
- C12Q1/6855Primary
Ligating adaptors · CPC title
incorporating an adaptor · CPC title
Bisulfite(s) · CPC title
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- What does patent US12385085B2 cover?
- The invention relates to a method of preparing and using a library of template polynucleotides suitable for use as templates in solid-phase nucleic acid amplification and sequencing reactions to determine the methylation status of the cytosine bases in the library. In particular, the invention relates to a method of preparing and analysing a library of template polynucleotides suitable for meth…
- Who is the assignee on this patent?
- Illumina Cambridge Ltd, Massachusetts Inst Technology
- What technology area does this patent fall under?
- Primary CPC classification C12Q1/6855. Mapped technology areas include Chemistry & Metallurgy.
- When was this patent published?
- Publication date Tue Aug 12 2025 00:00:00 GMT+0000 (Coordinated Universal Time) (B2). Legal status and post-grant events are not shown on this page.
- What related patents are in patentsdb?
- We list 2 related publications on this page (citations in our corpus or others sharing the same primary CPC).