Institute of Cell Biology and Genetics, College of Life Sciences, Zhejiang University, Zijingang Campus, Hangzhou, Zhejiang, 310058, People's Republic of China.
Mol Biotechnol. 2009 Nov;43(3):257-63. doi: 10.1007/s12033-009-9198-0. Epub 2009 Aug 11.
Here we present a novel method termed full-length normalization subtractive hybridization (FNSH) for efficiently generating subtracted cDNA libraries with a high degree of productivity. This method has the ability to isolate full-length differentially expressed genes from target samples. Normalization and subtraction of FNSH are performed simultaneously with efficiency equal to or even higher than that of suppression subtractive hybridization. Using FNSH, we have isolated at least 40 unique cDNAs that are expressed in terminal ampullae but not in the ovaries of the prawn Macrobrachium rosenbergii from 120 randomly picked subtracted clones. Sequence analysis shows that 37 of the 40 cDNAs are full length.
在这里,我们提出了一种新的方法,称为全长归一化消减杂交(FNSH),用于高效地生成具有高生产力的消减 cDNA 文库。该方法能够从目标样品中分离全长差异表达基因。FNSH 的归一化和消减是同时进行的,效率与抑制消减杂交相当,甚至更高。使用 FNSH,我们从 120 个随机挑选的消减克隆中分离出至少 40 个在对虾 Macrobrachium rosenbergii 的端囊而非卵巢中表达的独特 cDNA。序列分析表明,40 个 cDNA 中有 37 个是全长的。