Synthetic nucleic acid molecules are provided which are called indexing linkers and which can be selectively linked to unknown, or non-identical cohesive ends of nucleic acid fragments. Such indexing linkers are useful for the selective isolation, identification, amplification, labelling, and modification, of nucleic acid fragments, especially subsets of such fragments released by cleavage using Type IIS restriction endonucleases or restriction endonucleases recognizing interrupted palindromic sequences. Sets of such indexing linker molecules are described which are useful for indexing fragments released by cleavage of a genome. Methods are described for distinguishing such fragments using said indexing linker molecules for effecting detection, or further modification of such fragments; and for amplifying such fragments by the polymerase chain reaction.
This application is a Continuation of application Ser. No. 08/055,260, filed on May 3, 1993, now abandoned, which is a continuation of application Ser. No. 07/505,884, filed on Apr. 6, 1990, now abandoned.
This invention relates to a method for classifying (indexing) cDNA which has been reverse-transcribed from tissue- or cell-derived RNA, or DNA in a short period without duplication by using class-IIS restriction enzymes or a combination of a class-IIS and a class-II restriction enzymes. According to this invention, it is possible to analyse and diagnose variations such as tumors easily, correctly and promptly by comparing the analyzed pattern of genes expressed in a cell or tissue sample with the analyzed pattern of normal genes. This method is also applicable to the search and isolation of genes of physiologically active substances that are potential pharmaceuticals or causative genes of hereditary diseases, as well as the isolation of those genes that are useful for improving agricultural products.
Serial analysis of gene expression, SAGE, a method for the rapid quantitative and qualitative analysis of transcripts is provided. Short defined sequence tags corresponding to expressed genes are isolated and analyzed. Sequencing of over 1,000 defined tags in a short period of time (e.g., hours) reveals a gene expression pattern characteristic of the function of a cell or tissue. Moreover, SAGE is useful as a gene discovery tool for the identification and isolation of novel sequence tags corresponding to novel transcripts and genes.
Serial analysis of gene expression, SAGE, a method for the rapid quantitative and qualitative analysis of transcripts is provided. Short defined sequence tags corresponding to expressed genes are isolated and analyzed. Sequencing of over 1,000 defined tags in a short period of time (e.g., hours) reveals a gene expression pattern characteristic of the function of a cell or tissue. Moreover, SAGE is useful as a gene discovery tool for the identification and isolation of novel sequence tags corresponding to novel transcripts and genes.
A process for the categorization of nucleic acid sequences in which said sequences are linked to a population of adaptor molecules each exhibiting specificity for linking to a sequence including a predetermined nucleotide base, categorization of the resulting linked sequences being based upon selection for the base.
The present invention relates to novel methods for sequencing and mapping genetic markers in polynucleotide sequences using Type-IIs restriction endonucleases. The methods herein described result in the "capturing" and determination of specific oligonucleotide sequences located adjacent to Type-IIs restriction sites. The resulting sequences are useful as effective markers for use in genetic mapping, screening and manipulation.
