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US 8,906,617 B2Granted
Methods for sequencing individual nucleic acids under tension
AssigneeLife Technologies Corporation
InventorsJonathan M. Rothberg, John H. Leamon, John F. Davidson, Antoine M. Van Oijen, Wolfgang Hinz, Melville Davey, Bradley Hann, Jonathan Schultz
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Abstract
The invention provides apparatuses and methods of use thereof for sequencing nucleic acids subjected to a force, and thus considered under tension. The methods may employ but are not dependent upon incorporation of extrinsically detectably labeled nucleotides.
Claims16
A method for sequencing comprising:
- immobilizing a single stranded nucleic acid target by its 3′ end or 5′ end to a surface of a substrate, the single-stranded nucleic acid target including a plurality of tandem repeat nucleic acid sequences, the single-stranded nucleic acid target including a detectable moiety at its free end;
- applying at least one oligonucleotide complementary to at least a portion of a tandem repeat single stranded nucleic acid sequence of the nucleic acid target, the at least one oligonucleotide having a known sequence and hybridizing to the at least a portion of the tandem repeat nucleic acid sequence of the single stranded nucleic acid target;
- applying a force to the single stranded nucleic acid target;
- determining with an ion sensitive field effect transistor array and when the force is applied, a change in a length of the single stranded nucleic acid target arising from hybridizing the at least one oligonucleotide, the determining based on a change in a position of the detectable moiety caused by the conversion of single stranded bases to double stranded bases;
- and
- associating the change in the length of the single stranded nucleic acid target with a nucleotide sequence for the nucleic acid target.
3 more elements
A method for sequencing comprising:
- immobilizing a single stranded nucleic acid target by its 3′ end or 5′ end to a surface of a substrate, the single stranded nucleic acid target including a plurality of tandem repeat nucleic acid sequences, the single stranded nucleic acid target including a detectable moiety at its free end;
- applying a plurality of oligonucleotides complementary to at least a portion of the tandem repeat nucleic acid sequences, the plurality of oligonucleotides having a known sequencing and hybridizing to the plurality of tandem repeat nucleic acid sequences;
- applying a force to the single stranded nucleic acid target;
- determining with an ion sensitive field effect transistor array and when the force is applied, a change in a position of the detectable moiety arising from the hybridizing of the plurality of oligonucleotides and determining a change in a length of the single stranded nucleic acid target caused by the conversion of single stranded bases to double stranded bases;
- and
- associating the change in the length of the single stranded nucleic acid target with a nucleotide sequence for the single stranded nucleic acid target.
3 more elements
Dependencies are inferred from the claim text.