Abstract: A method is provided for adding a terminal sequence tag to nucleic acid molecules for use in RNA or DNA amplification. The method involves contacting with a mixture of oligonucleotides, each having a sequence tag template, a random sequence and a blocked 3? terminus, under conditions such that, the random sequence anneals with the nucleic acid molecules and the nucleic acid molecules are extended using the sequence tag template as template. For synthesis of RNA from DNA molecules having terminal sequence tags, the method includes forming DNA templates having a double stranded promoter sequence and synthesizing RNA from the DNA templates. For amplification of sequences from DNA molecules having terminal sequence tags, the method includes forming DNA templates by extension of one primer having a sequence that is complementary to the terminal sequence tag and another primer having a sequence that is derived form one of the DNA molecules.

Abstract: This invention relates to the use of a eukaryotic or prokaryotic DNA-directed RNA polymerase of a class that synthesizes cellular RNA, in a process for the amplification of a specific nucleic acid sequence or of its complement. It also relates to a new process for amplifying a specific nucleic acid sequence. The process includes one or more reactions which may take place in a single reaction vessel. In one instance, in a first reaction, the process includes providing a first RNA polymerase which uses a DNA first template to synthesize an RNA first template, and, in a second reaction, providing the RNA first template and a number of other reagents such that the reagents use the RNA first template to synthesize a DNA second template and an RNA second template. Thereafter a cycle ensues in which the reagents use the RNA second template to synthesize a DNA third template and multiple copies of the RNA second template. The RNA second template is the specific nucleic acid sequence or its complement.

Abstract: This invention relates to a process for amplifying a specific nucleic acid sequence or its complement at a relatively constant temperature and without serial addition of reagents. The process provides in a single reaction medium an RNA polymerase, DNA polymerase, a ribonuclease that hydrolyses RNA of an RNA-DNA hybrid without hydrolysing single or double-stranded RNA or DNA, and ribonucleoside and deoxyribonucleoside triphosphates. The process then provides an RNA first template in the reaction medium. The RNA first template comprises a sequence complementary to a specific nucleic acid sequence, minus-sense sequences for a promoter and initiation site that are recognized by the RNA polymerase, and a 5'-terminal sequence that is complementary to at least the minus-sense sequence of the initiation site. Thus, the RNA first template has an inverted repeat sequence which could fold into a 5'-terminal stem-loop structure.

Abstract: This invention relates to an improved process for amplifying a specific nucleic acid sequence. The process involves synthesizing single-stranded RNA, single-stranded DNA and Double-stranded DNA. The single-stranded RNA is a first template for a first primer, the single-stranded DNA is a second template for a second primer, and the double stranded DNA is a third template for synthesis of a plurality of copies of the first template. A sequence of the first primer or the second primer is complementary to a sequence of the specific nucleic acid and a sequence of the first primer or the second primer is homologous to a sequence of the specific nucleic acid. The improvement of the amplification process involves the addition of DMSO alone or in combination with BSA, which improves the specificity and efficiency of the amplification.