Linked dual barcode insertion constructs

Abstract

Contemporary gene sequencing techniques, including 'Next Generation Sequencing' techniques, can include sequencing a plurality of fragments of a target polynucleotide. However, tire limitations of existing sequencing techniques, and the often repetitive or otherwise difficult-to-sequence structure of natural polynucleotides, means that it can be difficult and/or expensive to generate accurate sequences. Methods provided herein include inserting dual polynucleotide 'barcodes,' along with neighboring primer sequences, into a target polynucleotide prior to other sequencing processes. These inserted barcodes can improve the accuracy of sequences generated for the target by adding ‘noise’ into the target, allowing subsequent sequencing techniques (e.g., alignment, stitching, etc.) to more accurately estimate the target-plus-barcodes sequence. The primers can cause the fragments to begin at points within the target that correspond to the beginning of other sequences, facilitating the stitching of sequence ends together. The barcodes can then be removed to provide the sequence of the target polynucleotide.