DETECTION OF GENETIC SEQUENCES USING PNA PROBES AND ISOTACHOPHORESIS

摘要

The present invention provides a system including: a peptide nucleic acid (PNA) molecule; a DNA molecule, an RNA molecule, or a combination thereof; and an Isotachophoresis (ITP) system. Furthermore, the invention provides a method for sequence-specifically separating and/or identifying a nucleic acid molecule of interest by utilizing the system of the invention to separate and possibly label and/or detect a nucleic acid molecule of interest.

主权项

1. A method for sequence-specifically detecting and/or separating a nucleic acid molecule, comprising the steps of:
a. contacting a mixture of nucleic acid molecules with a labeled peptide nucleic acid molecule (PNA) having a sequence of interest in a first solution and obtaining a nucleic acid molecule/PNA hybrid; b. introducing said first solution between a fast leading electrolyte and a slow terminating electrolyte of an isotachophoresis (ITP) system, said ITP system comprises a second solution of high effective mobility leading electrolyte (LE) ions and a third solution of low effective mobility trailing electrolyte (TE); c. applying an electric field across said second solution and said third solution; wherein said nucleic acid molecule/PNA hybrid but not free PNA focus at the sharp LE-TE interface in said ITP system, wherein said TE has a higher mobility than the PNA probe and said TE has a lower mobility than the nucleic acid molecule/PNA hybrid, thereby sequence-specifically separating a nucleic acid molecule.