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In-Gel Purified Ditags Direct Synthesis Of Highly Efficient Sage Libraries, Saroj P. Mathupala, Andrew E. Sloan
In-Gel Purified Ditags Direct Synthesis Of Highly Efficient Sage Libraries, Saroj P. Mathupala, Andrew E. Sloan
Wayne State University Associated BioMed Central Scholarship
Abstract
Background
SAGE (serial analysis of gene expression) is a recently developed technique for systematic analysis of eukaryotic transcriptomes. The most critical step in the SAGE method is large scale amplification of ditags which are then are concatemerized for the construction of representative SAGE libraries. Here, we report a protocol for purifying these ditags via an 'in situ' PAGE purification method. This generates ditags free of linker contaminations, making library construction simpler and more efficient.
Results
Ditags used to generate SAGE libraries were demarcated 'in situ' on preparative polyacrylamide gels using XC and BPB dyes, which precisely straddle the ditag …