cDNA Microarray: Gene Expression Profiling of Small Cell Populations
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| Year of publication | 2008 |
| Type | Conference abstract |
| MU Faculty or unit | |
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| Description | cDNA microarray technology is widely used in various biological and medical disciplines to determine gene expression profiles. Unfortunately, this technology requires a large quantity of input RNA. Since there is an increasing need to analyze more precisely defined cell sub-populations with low cell counts, working protocols using a minimal number of input cells are needed. Optimal RNA isolation and its accurate amplification are crucial to the success of these protocols. The goal was to find the best method of isolating and amplifying RNA from a small number of cells. We used the HL60 cell line in the search for a suitable protocol that could be applied to clinical samples of CD34+ hematopoietic cells. These cells make up only a very small population in bone marrow (1- 4% of all cells). Our evaluation of various methods and kits available in the market revealed that the combination of RNAqueous Kit (Ambion, USA) for RNA isolation and the SenseAmp Plus Kit (Genisphere, USA) for linear one-round RNA amplification produced the best results. The agreement of the expression ratio for genes detected in unamplified RNA and amplified RNA was 97.6 %. This verified protocol describes a reliable and reproducible method for producing enough input RNA for microarray experiments when the number of cells is limited. |
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