| 主权项 |
1. A method of making a dual barcoded nucleic acid molecule comprising:
(a) coupling one strand of a stem-loop oligonucleotide to at least one end of a nucleic acid molecule to form a first barcode-coupled nucleic acid molecule, wherein the stem-loop oligonucleotide includes an intramolecular inverted repeat and a loop, and wherein the inverted repeat includes the first barcode; (b) displacing one strand of the stem-loop oligonucleotide from the first barcode-coupled nucleic acid molecule by strand displacement or by nick translation polymerization to form a first barcoded nucleic acid molecule; (c) annealing a primer to the first barcoded nucleic acid molecule, wherein the primer comprises a first portion complementary to the first barcoded nucleic acid molecule and a second portion comprising a second barcode; and (d) extending the annealed primer to form a dual barcoded nucleic acid molecule, the dual barcoded nucleic acid molecule comprising the second barcode, the first barcode, and at least a portion of the nucleic acid molecule. |
