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I-SAGE™Long Kit

Improved Method for Serial Analysis of Gene Expression

LongSAGE™ (serial analysis of gene expression) is an adaptation of SAGE™ technology. Like SAGE™(1), LongSAGE™(2) allows comprehensive, digital genome-wide expression analysis. The expression profiles generated are sensitive enough to detect low-abundance transcripts, making it possible to identify novel genes without prior sequence knowledge. LongSAGE™ introduces significant advantages over the traditional SAGE™ method:

  • The increased length of the LongSAGE™ (21 bp) "tag" provides greater specificity, making discrimination between genes and the detection of novel genes easier.
  • The additional tag length enables the mapping of the "tags" to their genomic location, facilitating cloning and subsequent analysis.
The I-SAGE™ Long Kit uses an improved protocol that makes LongSAGE™ library construction more robust.

How the I-SAGE™ Long Kit Works

  • I-SAGE™ Long Kit protocol uses standard molecular biology techniques to construct LongSAGE™ libraries. Libraries are constructed by isolating a defined 21 bp region called a "tag", rather than the entire sequence of each transcript. The tag contains enough sequence to uniquely identify each mRNA transcript. These tags are ligated into concatemers, cloned, and sequenced (Figure 1). There is no PCR size bias because all templates for PCR are 130 bp. Rapid high-throughput sequencing of the concatemer clones identifies the number of individual tags in each clone. SAGE™ analysis software digitally archives the sequencing data for immediate and future analysis. SAGE™ data can be used to search the SAGEmap database(3) (http://www.ncbi.nlm.nih.gov/SAGE) or SAGE Genie(4) (http://cgap.nci.nih.gov/SAGE) reference databases and compared to other SAGE™ libraries.

Important Licensing Information

  • These products may be covered by one or more Limited Use Label Licenses (See the Invitrogen catalog or our web site, http://www.invitrogen.com). By the use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Products referred to herein which are the subject of one or more of U.S. Patent No. 5,910,438 and 6,180,407, and/or corresponding foreign patents, are sold under license from the Université Libre de Bruxelles for research purposes only. See also Limited Use Label License No. 54: ccdB-Fusion Vectors. SAGE™ and LongSAGE™ are trademarks of Genzyme Corporation. I-SAGE™ and I-SAGE™ Long are trademarks of Invitrogen Corporation.

References

    1. Velculescu, V.E. et al. (1995) Science 270: 484-487
    2. Saha, S. et al. (2002) Nature Biotechnology 19: 508-512
    3. Lash, A. et al. (2000) Genome Research 10: 1051-1060
    4. Boone, K. et al. (2002) PNAS 99: 11287-11292