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荧光通用引物差异显示PCR(FLUPD DD PCR)是以荧光通用引物和相应 1 0碱基随机引物进行DD PCR扩增 ,在荧光自动测序仪上进行电泳 .通过对电泳图谱的分析进行特异条带的确定 ,使准确度和自动化程度大大提高 ,尤其对于表达量上有差异的cDNA片段 ,这一方法比传统的DD PCR更为方便 .cy5标记的通用引物的使用和银染显带直接进行条带回收 ,进一步提高了该方法的效率和准确性 .应用这一方法 ,对血清饥饿处理的胶质瘤细胞的特异表达的mRNA进行了初步研究 ,并得到了 4条与该处理有关的EST .经过与GenBank数据库比较发现 ,其中 3条为未知的新EST ,1条与小鼠的EST完全同源 ,表明血清饥饿涉及到许多新基因的开启表达或某些基因的表达量上调 ,可诱发细胞发生多方面的生理反应 .
Fluorescent Universal Primer Differential Display PCR (FLUPD DD PCR) uses DD PCR amplification with fluorescent universal primers and the corresponding 10 base random primers, and performs electrophoresis on a fluorescent automatic sequencer. Specific banding is performed by analysis of the electrophoretic patterns. Determined to greatly improve the accuracy and automation, especially for cDNA fragments with different expression levels, this method is more convenient than traditional DD PCR. The use of cy5-labeled universal primers and silver staining bands are directly carried out The recovery has further improved the efficiency and accuracy of the method. Using this method, a preliminary study was conducted on mRNAs specifically expressed in serum-starved glioma cells, and four ESTs related to the treatment were obtained. Compared with the GenBank database, three of them are unknown new ESTs, and one is completely homologous to mouse ESTs, indicating that serum starvation involves the initiation of expression of many new genes or the up-regulation of the expression of certain genes, which can induce cell development. A variety of physiological reactions.