 | TagSorterTagSorter is a database program built to simplify the collation, sorting, comparison and identification of SAGE tag sequences |
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TagSorter Description
TagSorter is a database program built to simplify the collation, sorting, comparison and identification of SAGE tag sequences. Serial Analysis of Gene Expression (SAGE) is a high through put technique for measuring gene expression profiles (Velculescu et al, 1995). It reveals both the identity of transcripts expressed and their relative abundance. It can also be used as a means of gene discovery. Use of SAGE has steadily grown since its invention in 1995 and gene expression profiles have been generated from a variety of tissues and organisms. A brief overview of the SAGE technique is as follows. Messenger RNA is harvested from the experimental tissue and is converted to double stranded cDNA in such a way that the polyA tail of each message is biotin labeled. This cDNA is digested with the restriction enzyme NlaIII and using the biotin label the 3'-most NlaIII digestion products are captured. A BsmFI restriction site adapter is then ligated to these fragments. BsmFI cleaves DNA at a position approximately 14 base pair from its recognition site. This results in short expressed sequence tags (EST) from a defined position within each of the mRNA transcripts present in the original tissue. These fragments are then ligated together to form a concatemer of EST's which are cloned into a standard plasmid vector and sequenced. Each clone can contain identifying tags for up to sixty mRNA transcripts. Since the relative position of each tag is known and they consist of 10 base pairs of novel sequence, once the NlaIII site is excluded, that uniquely identify the mRNA transcript from which they were generated.
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