HIGH THROUGHPUT SEQUENCING OF END REGIONS OF LONG LINEAR DNAs

摘要

This invention relates to linking, amplifying and sequencing of two ends of long linear DNAs. In particular, this invention provides methods for pairing and sequencing VH and VL genes that encode two parts of one immunoglobulin. The method of the present invention can be applied to rapid antibody discovery and engineering.

主权项

1. A method for obtaining sequence identities of a plurality of clones of a phage-displayed antibody library, comprising,
providing plurality of linear double stranded DNA (dsDNA) molecules from said clones, each dsDNA molecule containing (1) a 5′ end segment at the 5′ end of the dsDNA molecule and (2) a 3′ end segment at the 3′ end of the dsDNA molecule, the two end segments being separated by a spacer segment; fusing the two end segments to form a circular dsDNA molecule and a joined section on the dsDNA molecule that has the two end segments joined by a junction, and sequencing the joined section of each dsDNA molecule from each clone using high throughput sequencing thereby obtaining the sequence identities of the plurality of clones.