Barcoded Solid Supports and Methods of Making and Using Same

Abstract

Aspects of the present disclosure include methods of making barcoded solid supports. In some embodiments, the methods include producing a concatemer by rolling circle amplification (RCA) of a circular nucleic acid template, where the circular nucleic acid template includes a barcode and a stem-loop forming region, and where the concatemer includes a plurality of linked units, each unit including the barcode and a stem-loop structure formed from the stem-loop forming region. Such methods further include disposing the concatemer on a solid support to produce a barcoded solid support including a plurality of the stem-loop structures extending from the surface of the solid support. The methods may further include treating the stem-loop structures with an agent that produces stem structures having ends compatible with target nucleic acids, and attaching the target nucleic acids to the stem structures. Barcoded solid supports and methods of using the barcoded solid supports are also provided.

1 .- 17 . (canceled) 
     
     
         18 . A barcoded solid support, comprising:
 a concatemer disposed on a solid support, wherein the concatemer comprises a plurality of linked units, each unit comprising a barcode and a stem-loop structure, and wherein a plurality of the stem-loop structures extend from the surface of the solid support.   
     
     
         19 . The barcoded solid support of  claim 18 , wherein:
 the concatemer comprises a plurality of first binding members;   the solid support is functionalized with a plurality of second binding members having affinity for the first binding members; and   the first binding members are bound to the second binding members.   
     
     
         20 . The barcoded solid support of  claim 19 , wherein the first binding members are non-covalently bound to the second binding members. 
     
     
         21 . The barcoded solid support of  claim 20 , wherein the first binding members comprise biotin and the second binding members comprise streptavidin, avidin, or anti-biotin antibodies. 
     
     
         22 . The barcoded solid support of  claim 19 , wherein the first binding members are enriched in regions between the stem-loop structures. 
     
     
         23 . The barcoded solid support of  claim 18 , wherein each of the plurality of linked units of the concatemer further comprises a partial or complete sequencing adapter. 
     
     
         24 . The barcoded solid support of  claim 18 , wherein the solid support comprises a bead. 
     
     
         25 .- 29 . (canceled)