Method to produce dna molecules with repeating units for use in single-molecule assays
AssigneeLumicks Dsm Holding B.V.
InventorMarco SIMONETTA
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Abstract
The invention relates to a method for producing linear DNA molecules with at least two or more repeating units of a target DNA sequence for use as DNA substrate in single-molecule assays, wherein a target DNA is subjected to a ligation reaction and the ligation reaction is stopped when the reaction reaches the end of the linear phase to block circularization of the DNA molecules. Compositions comprising such linear DNA molecules and use of such compositions in single-molecule assays are also provided.
Claims20
A method for producing linear DNA molecules with two or more repeating units of a target DNA sequence, the method comprising steps of:
- a) providing a plurality of the target DNA sequence and a DNA ligase and subjecting the plurality of target DNA sequence to a ligation reaction to obtain a first mixture of DNA molecules varying in length and number of repeats, wherein each DNA molecule in the first mixture of DNA molecules comprises at least two or more repeating units of the target DNA sequence, and
- b) stopping the ligation reaction at an end of a linear phase of the ligation reaction, and
- c) recovering said first mixture of DNA molecules.
A method for producing linear DNA molecules with two or more repeating units of a target DNA sequence, the method comprising steps of:
- a) providing a plurality of the target DNA sequence and a DNA ligase and subjecting the target DNA sequence to a ligation reaction to obtain a first mixture of DNA molecules varying in length and number of repeats, wherein each DNA molecule in the first mixture comprises at least two or more repeating units of the target DNA sequence,
- b) adding DNA adaptor molecules to the first mixture of DNA molecules at an end of a linear phase of the ligation reaction to obtain a second mixture of DNA molecules varying in length and number of repeats, wherein each DNA molecule in the second mixture of DNA molecules comprises at least two or more repeating units of the target DNA sequence, and part of the DNA molecules in the second mixture consists of adapted linear DNA molecules.
A composition comprising a mixture of DNA molecules varying in length and number of repeats, wherein each DNA molecule comprises at least two or more repeating units of a target DNA sequence, and part of the DNA molecules consists of functionalized linear DNA molecules.
Dependencies are inferred from the claim text.