The purpose of nucleic acid sequencing is to determine the order of the bases in DNA or RNA molecules. In the past decades, both chemical degradation and enzymatic (dideoxy) methods have been serving as the working horses for nucleic acid sequencing. However, in these days of more advanced technology, the development of high-throughput approaches makes it more convenient to contract sequencing jobs. In the past few years, nucleic acid sequencing throughput has increased over 50-fold since it has undergone significant advancement from the discovery of the Sanger’s approach to the so-called “second-generation” sequencing methods, such as pyrosequencing, Solexa sequencing, and SOLiD sequencing. Moreover, the so-called “third-generation technology”, a gel-free method incorporating properties such as enhanced fluorescent detection of individual labeled bases in flow cytometry and direct reading of the strand, has also been invented. Some selected uses of nucleic acid sequencing include the study of the structure and organization of genes and the elucidation of gene regulation mechanisms. In this review, we aim to describe recent patents for nucleic acid sequencing and its applications by dividing them into three categories: 1) chain elongation and termination methods; 2) nanopore sequencing methods; and 3) other approaches and applications. In addition, the perspectives of future developments in nucleic acid sequencing are also provided at the end of this review.