Base-detecting pore
US-9447152-B2 · Sep 20, 2016 · US
Method for attaching one or more polynucleotide binding proteins to a target polynucleotide
US11725235B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-11725235-B2 |
| Application number | US-201916540425-A |
| Country | US |
| Kind code | B2 |
| Filing date | Aug 14, 2019 |
| Priority date | Jan 22, 2014 |
| Publication date | Aug 15, 2023 |
| Grant date | Aug 15, 2023 |
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- Title
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Abstract
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The invention relates to new methods of attaching one or more polynucleotide binding proteins to a target polynucleotide. The invention also related to new methods of characterising target polynucleotides.
First claim
Opening claim text (preview).
The invention claimed is: 1. A method for attaching one or more polynucleotide binding proteins to a target polynucleotide, comprising: (a) providing a complex formed by the one or more polynucleotide binding proteins and one or more loading moieties comprising a loading polynucleotide, wherein the one or more polynucleotide binding proteins are bound to the loading polynucleotide; and (b) attaching the loading polynucleotide of the one or more loading moieties of the complex to the target polynucleotide, wherein the one or more polynucleotide binding proteins of the complex retain their function and ability to bind to the loading polynucleotide of the one or more loading moieties after said attaching the loading polynucleotide of the one or more loading moieties of the complex to the target polynucleotide; and wherein the one or more polynucleotide binding proteins are one or more helicases, and wherein the one or more helicases are He1308 helicases, RecD helicases, XPD helicases or Dda helicases. 2. A method for attaching one or more polynucleotide binding proteins to a target polynucleotide, comprising: (a) providing a complex formed by the one or more polynucleotide binding proteins and one or more loading moieties comprising a loading polynucleotide, wherein the one or more polynucleotide binding proteins are bound to the loading polynucleotide; and (b) attaching the loading polynucleotide of the one or more loading moieties of the complex to the target polynucleotide, wherein the one or more polynucleotide binding proteins of the complex retain their function and ability to bind to the loading polynucleotide of the one or more loading moieties after said attaching the loading polynucleotide of the one or more loading moieties of the complex to the target polynucleotide; and wherein the one or more polynucleotide binding proteins are derived from helicases and are stalled at one or more spacers on the loading polynucleotide. 3. The method according to claim 1 or 2 , wherein the method further comprises binding the polynucleotide binding proteins to the one or more loading moieties before step (a). 4. The method according to claim 1 or 2 , wherein the one or more polynucleotide binding proteins are two or more polynucleotide binding proteins. 5. The method according to claim 4 , wherein the two or more polynucleotide binding proteins are not directly attached to one another but are indirectly attached to one another via the one or more loading moieties. 6. The method according to claim 4 , wherein the two or more polynucleotide binding proteins are different from one another. 7. The method according to claim 1 or 2 , wherein the loading polynucleotide is a single-stranded polynucleotide. 8. The method according to claim 1 or 2 , wherein the loading polynucleotide is a Y adaptor and the Y adaptor comprises a leader sequence which preferentially threads into a pore. 9. The method according to claim 1 or 2 , wherein the loading polynucleotide is a bridging moiety, the bridging moiety is a hairpin loop and the stem of the hairpin loop is less than 200 nucleotide pairs in length.
Assignees
Inventors
Classifications
- C12Q1/6869Primary
Methods for sequencing · CPC title
Ligases (6) · CPC title
involving nucleic acids · CPC title
characterised by the detection means (C12Q1/6804 takes precedence) · CPC title
Enzymatic or biochemical coupling of nucleic acids to a solid phase · CPC title
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Frequently asked questions
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- What does patent US11725235B2 cover?
- The invention relates to new methods of attaching one or more polynucleotide binding proteins to a target polynucleotide. The invention also related to new methods of characterising target polynucleotides.
- Who is the assignee on this patent?
- Oxford Nanopore Tech Ltd, Oxford Nanopore Tech Plc
- What technology area does this patent fall under?
- Primary CPC classification C12Q1/6869. Mapped technology areas include Chemistry & Metallurgy.
- When was this patent published?
- Publication date Tue Aug 15 2023 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 12 related publications on this page (citations in our corpus or others sharing the same primary CPC).