Shi Xin, Gao Nai-Rong, Hu Hao-Lin, Huo Ming-Dong, Tang Wen-Hao
Department of General Surgery, Zhongda Hospital, Southeast University, Nanjing 210009, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2003 Aug;19(3):310-2.
To investigate the method of detecting gene expression in colon tissue at a single cell level.
Individual cell(s) were picked up from colon frozen section using laser microdissection. RNA was extracted, reverse transcribed to complementary DNA (cDNA). cDNA was then analyzed by nested reverse transcription polymerase chain reaction (nested RT-PCR) using two pairs of primers.
Single cell(s) were selectively picked up using an ultraviolet laser micromanipulator. RNA was extracted, reverse transcribed and used for nested RT-PCR. Amplification products of cDNA from down to a single cell could be clearly visualized in the agarose gel.
The combined utilization of laser microdissection and nested RT-PCR provides an opportunity to analyze gene expression at single cell(s) level in colon tissue.
研究在单细胞水平检测结肠组织中基因表达的方法。
使用激光显微切割技术从结肠冰冻切片中挑选单个细胞。提取RNA，逆转录为互补DNA（cDNA）。然后使用两对引物通过巢式逆转录聚合酶链反应（巢式RT-PCR）分析cDNA。
使用紫外激光显微操作仪选择性地挑选单个细胞。提取RNA，进行逆转录并用于巢式RT-PCR。在琼脂糖凝胶中可以清晰地看到从单个细胞获得的cDNA扩增产物。
激光显微切割技术与巢式RT-PCR的联合应用为分析结肠组织中单个细胞水平的基因表达提供了机会。
