Chapter title |
Laser Capture Microdissection and Isolation of High-Quality RNA from Frozen Endometrial Tissue
|
---|---|
Chapter number | 8 |
Book title |
Laser Capture Microdissection
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7558-7_8 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7557-0, 978-1-4939-7558-7
|
Authors |
Michele Cummings, Georgia Mappa, Nicolas M. Orsi, Cummings, Michele, Mappa, Georgia, Orsi, Nicolas M. |
Abstract |
Laser capture microdissection (LCM) allows expression profiling of specific cell populations within tissues. However, isolation of high-quality RNA from laser capture microdissected frozen tissue is beset by problems arising from intrinsic tissue RNase activity. Herein, we describe an optimized staining/LCM/RNA extraction protocol developed for the isolation of epithelial RNA from frozen tissue sections using human endometrial cancer as a model tissue. This method combines excellent, reproducible visualization of tissue morphology with the isolation of high-integrity RNA suitable for downstream applications such as expression microarray analysis. We present quantitative and qualitative RNA data obtained from >200 endometrial epithelial samples (normal, hyperplastic, and cancerous), where 92% of samples had RIN values of 7 and above and highlight common pitfalls faced by investigators. This method should also be broadly applicable to a range of other tissue types. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 7 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Master | 2 | 29% |
Professor > Associate Professor | 1 | 14% |
Student > Ph. D. Student | 1 | 14% |
Other | 1 | 14% |
Researcher | 1 | 14% |
Other | 0 | 0% |
Unknown | 1 | 14% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 3 | 43% |
Neuroscience | 2 | 29% |
Unknown | 2 | 29% |