Sequence of surface immobilized polymers utilizing microfluorescence detection

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.

1 . 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.  
     
     
         2 . A method of  claim 1 , wherein said polymers are polynucleotides.  
     
     
         3 . A method of  claim 1 , wherein the labeling of the terminal marker comprises incorporation of a labeled terminal monomer selected from the group of nucleotides consisting of adenine, cytidine, guanidine and thymidine.  
     
     
         4 . 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.  
     
     
         5 . A method of  claim 4 , wherein said solution comprises a reagent which marks said positionally distinct polymer with a fluorescent label moiety.  
     
     
         6 . A method of  claim 4 , wherein said terminal subunit is selected from the group consisting of adenosine, cytosine, guanosine, and thymine.  
     
     
         7 . 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.  
     
     
         8 . A method of  claim 7 , wherein said oligonucleotide subunit is a single nucleotide.  
     
     
         9 . A method of  claim 8 , wherein the marking comprises elongating said primer with a labeled nucleotide which is complementary to a template.  
     
     
         10 . A method of  claim 7 , wherein the marking step uses a polymerase and a blocked and labeled adenine.  
     
     
         11 - 33 . (canceled)