Synthetic long read dna sequencing

Abstract

The disclosure describes a method for sequencing long portions of DNA sequence by assembling a plurality of shorter polynucleotide reads. Generally, the method includes annealing a plurality of primers to a denatured DNA molecule, appending a barcode polynucleotide to the 5′ end of the primer, subjecting the DNA molecules to a plurality of cycles of (1) pooling, (2) dividing, and (3) appending a barcode polynucleotide to the 5′ end of the primer, sequencing the barcode polynucleotides and the genomic DNA, and assembling the short read polynucleotide sequences having identical barcode polynucleotides.

1 - 12 . (canceled) 
     
     
         13 . A polynucleotide comprising:
 a 5′ region comprising a randomized sequence of nucleotides;   an amplification sequence 3′ to the 5′ region that is complementary to an amplification primer; and   a 3′ region 3′ to the amplification sequence.   
     
     
         14 . The polynucleotide of  claim 13 , wherein the 5′ region comprises from five to 25 nucleotides. 
     
     
         15 . The polynucleotide of  claim 13 , wherein the amplification sequence comprises from six to 50 nucleotides. 
     
     
         16 . The polynucleotide of  claim 13 , wherein the 3′ region comprises from one to 10 nucleotides.