151. [Application of microdissection combined with RNA linear amplification].
- Author
-
Han WN, Li H, Wang S, Zhou W, and Yao KT
- Subjects
- Dissection methods, Humans, Nasopharyngeal Neoplasms pathology, RNA, Neoplasm isolation & purification, RNA, Neoplasm metabolism, Reverse Transcriptase Polymerase Chain Reaction, Nasopharyngeal Neoplasms genetics, RNA, Neoplasm genetics
- Abstract
Background & Objective: Microdissection has become indispensable for the selective analysis of stroma-free tumor cell. However, To obtain sufficient RNA from microdissected cells is difficult. The study was designed to seek a specific way to separate nasopharyngeal carcinoma (NPC) cells from stromal cells and to amplify the RNA from microdissected NPC cells., Methods: NPC cells were obtained using microdissection from frozen NPC tissue sections, then RNA was extracted from the microdissected NPC cells and reverse transcribed in vitro. The expression levels of beta-actin and GADPH in amplified RNA were detected using RT-PCR., Results: About 20,000-40,000 NPC cells were obtained, RNA was extracted from the cells, about 0.5-2.5 kb RNA fragments were obtained after RNA linear amplification and beta-actin and GADPH levels were integral., Conclusion: Microdissection combined with RNA linear amplification can be used to successfully obtain pure NPC cells, the integrity of amplified RNA is good and can be used in further research.
- Published
- 2002