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Laser Capture Microdissection and Isolation of High-Quality RNA from Frozen Endometrial Tissue
- Source :
- Methods in Molecular Biology ISBN: 9781493975570
- Publication Year :
- 2018
- Publisher :
- Springer New York, 2018.
-
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.
- Subjects :
- 0301 basic medicine
Frozen section procedure
Chemistry
Microarray analysis techniques
RNA
Endometrium
Molecular biology
Epithelium
Gene expression profiling
03 medical and health sciences
030104 developmental biology
medicine.anatomical_structure
medicine
RNA extraction
Laser capture microdissection
Subjects
Details
- ISBN :
- 978-1-4939-7557-0
- ISBNs :
- 9781493975570
- Database :
- OpenAIRE
- Journal :
- Methods in Molecular Biology ISBN: 9781493975570
- Accession number :
- edsair.doi...........6395a5068ef94cb320b266c75679ccdc