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Isolate and Analyze Total RNA from Cells Harvested by Laser Capture ,,, Microdissection

enes RT-PCR Primers for Human Gene Transcripts of Varying Abundance Levels according to the instructions.3 RNA of all abundance levels was detected in samples isolated from 200 cells. High abundance RNA was detected in a portion of the RT-PCR product equivalent to 2.5 cells, medium abundance RNA from a portion equivalent to 10 cells, and low abundance RNA from portions equivalent to 13 cells (Figure 1).

In addition, RNA isolated from malignant glioma cells using the StrataPrep microprep kit has been used in differential display experiments. These experiments identified several differentially expressed genes in the invasive rim of the tumor relative to the noninvasive tumor core.4

Conclusions

The StrataPrep total RNA microprep kit is ideal for the isolation of RNA from small cell samples harvested by LCM. The method generates pure, intact RNA with high efficiency that can be effectively amplified by conventional and quantitative RT-PCR.

Acknowledgments

The authors thank Michael Berens, Neurology Research, Barrow Neurological Institute, Phoenix, AZ for providing LCM samples.

REFERENCES

1. Emmert-Buck, M. R., et al. (1996) Science 274: 998-1001.
2. Dolter, K. and Braman J. (2000) Strategies 13: 56-58.
3. Rogers, B. and McKenzie, D. (1998) Strategies 11: 100-102.
4. Berens, M. E., et al. (2000) Proceedings of the American Association for Cancer Research 41:232, Abstract 1477.


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