Dna construct for sequencing and method for preparing the same

Abstract

A DNA construct comprising multiple units sequentially attached one to the other, wherein a unit comprises: a segment; an index attached to one end of the segment; an identifier attached to another end of the segment; an introducer attached to a 5′-end of either the index or the identifier, and a closure attached to a 5′-end of a remaining either identifier or index. A method for preparing the DNA construct and a method for analyzing a sequence of the DNA construct, as well as various embodiments thereof are disclosed herein.

1 . A DNA construct comprising multiple units sequentially attached one to the other, wherein a unit comprises:
 a segment;   an index attached to one end of the segment;   an identifier attached to another end of the segment;   an introducer attached to a 5′-end of either the index or the identifier, and   a closure attached to a 5′-end of a remaining either identifier or index.   
     
     
         2 . The DNA construct of  claim 1 , wherein the length of the DNA construct is at least substantially 1,000 bp. 
     
     
         3 . The DNA construct of  claim 1 , wherein the length of the segment is up to substantially 1,000 bp. 
     
     
         4 . The DNA construct of  claim 1 , wherein the length of the segment is in the range of substantially 100-500 bp. 
     
     
         5 . A method for preparing A DNA construct, the method comprising:
 obtaining a segment;   attaching an index, an identifier, an introducer and a closure to the segment,   wherein   the index attached to one end of the segment;   the identifier is attached to another end of the segment;   the introducer is attached to a 5′-end either the index or the identifier; and   the closure is attached to a 5′-end of a remaining either identifier or index, giving rise to a pre-mature unit;   amplifying the pre-mature unit with primers specific to the introducer and closure, giving rise to a double stranded mature unit;   phosphorylating 5′-ends of the strands of the mature unit, and sequentially attaching mature units.   
     
     
         6 . A method for analyzing a sequence of a DNA construct according to  claim 1 , the method comprising:
 separating sequences of units one from the other;   grouping units having the same index for obtaining same index groups;   grouping units having the same segment sequence in each same index group for obtaining same segment groups;   grouping units having the same identifier sequence in each same segment group for obtaining same identifier groups;   collapsing multiple segment sequences in each same identifier group to a single sequence that accurately represents the sequence of the target sequence according to which the segment was obtained.