Manual microdissection combined with antisense RNA-longSAGE for the analysis of limited cell numbers.
Establishing a gene expression profile of defined subtypes of cells within an organ is still challenging because it frequently requires microdissection and subsequent amplification of the limited amount of messenger RNA (mRNA) isolated from the microdissected tissue in order to be able to proceed with comprehensive gene expression analyses via microarray or serial analysis of gene expression (SAGE) technology. Here we describe a manual microdissection strategy for the isolation of high-quality RNA. Furthermore, a strategy for combining linear amplification of RNA with longSAGE is described that allows the use of antisense RNA (aRNA) generated via the well-established linear amplification of RNA procedure together with the conventional SAGE or longSAGE technology.
Lüttges
Jutta
J
Institute für Pathology, Saarbrücken Hospital, Saarbrücken, Germany.
Hahn
Stephan A
SA
Heidenblut
Anna M
AM
D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods*; D015723:Gene Library; D042282:Microdissection; D008957:Models, Genetic; D008967:Molecular Biology / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D012313:RNA / Q000737:chemistry; D016372:RNA, Antisense / Q000235:genetics*; D014169:Transformation, Bacterial
D018076:DNA, Complementary; D016372:RNA, Antisense; D012313:RNA