Sequencing of surface immobilized polymers utilizing microfluorescence detection
OwnerWilliam J Dower, Stephen P.A. Fodorinventors
Life of this patent
Estimating the term…
Abstract
Means for simultaneous parallel sequence analysis of a large number of biological polymer macromolecules. Apparatus and methods may use fluorescent labels in repetitive chemistry to determine terminal manomers on solid phase immobilized polymers. Reagents which specifically recognize terminal manomers are used to label polymers at defined positions on a solid substrate.
Claims33
A method for determining the positions of polymers which terminate with a given monomer, where said polymers are attached to a surface having a plurality of positionally distinct polymers attached thereto, said method comprising the steps of:
- labeling a terminal monomer in a monomer type specific manner;
- and
- scanning said surface, thereby determining the positions of said label.
A method for concurrently determining which subset of a plurality of positionally distinct polymers attached to a solid substrate at separable locations terminates with a given terminal subunit, said method comprising the steps of:
- mixing said solid substrate with a solution comprising a reagent, which selectively marks positionally distinct polymers which terminate with said given terminal subunit;
- and
- determining with a detector which separable locations are marked, thereby determining which subset of said positionally distinct polymers terminated with said given terminal subunit.
A method for determining which subset of a plurality of primer polynucleotides have a predetermined oligonucleotide, wherein the polynucleotides are complementary to distinctly positioned template strands which are attached to a solid substrate, said method comprising the steps of:
- selectively marking said subset of primer polynucleotides having the predetermined oligonucleotide;
- and
- detecting which polynucleotides are marked.
A method for concurrently obtaining sequence information on a plurality of polynucleotides by use of a single label detector, said method comprising the steps of:
- attaching a plurality of positionally distinct polynucleotides to a solid substrate at separable locations;
- labeling said plurality of polynucleotides with a terminal nucleotide specific reagent, said label being detectable using said label detector;
- determining whether said specific labeling reagent has labeled each separable location.
A method for concurrently obtaining sequence information on a plurality of polynucleotides, said method comprising the steps of:
- attaching distinct polynucleotides to a plurality of distinct solid substrates;
- labeling said plurality of solid substrates with a terminal nucleotide specific labeling reagent;
- and
- determining whether said specific labeling reagent has labeled each distinct substrate.
A method for simultaneously sequencing a plurality of polymers made up of monomer units, said plurality of polymers attached to a substrate at definable positions, said method comprising the steps of:
- mixing said substrate with a reagent which specifically recognizes a terminal monomer, thereby providing identification among various terminal monomer units;
- and
- scanning said substrate to distinguish signals at definable positions on said substrate;
- and
- correlating said signals at defined positions on said substrate to provide sequential series of sequence determinations.
A method for sequencing a plurality of distinctly positioned polynucleotides attached to a solid substrate comprising the steps of:
- hybridizing complementary primers to said plurality of polynucleotides;
- elongating a complementary primer hybridized to a polynucleotide by adding a single nucleotide;
- and
- identifying which of said complementary primers have incorporated said nucleotide.
Dependencies are inferred from the claim text.