- 作者: 歐樂君; 羅彩瑜; 王興華; 朱廣邦
- 作者服務機構: 臺北榮民總醫院醫學研究部; 臺北榮民總醫院分子醫學中心; 國立陽明大學醫事技術系
- 中文摘要:
當一段兩端為平頭或相同凸出的DNA接入載體時,其存在有兩個方向性,所以對於一個有陽性反應的純株,通常
需要再以限制?或鹼基定序方法確認其方向及接點的正確性,在此我們提出以跨越接合點,16-18個鹼基所組成的寡
核酸探針作雜交篩選,於是陽性純株,正確方向及接點鹼基序列無誤可在一次篩選中完成。
Y - 英文摘要: Cloning of DNA fragments with blunt ends or identical protruding ends results in colonies withthe insertion sequence existing in two possible orientations. The orientation and the junctionalsequence of the positive clones need to be established by means of restriction analysis and/or se-quencing. Here, we proposed a rapid one-step method for the screening of clones not only with thedesired orientation, but also with an intact junctional sequence. In this method, a 16-18 meric oligo-nucleotide probe synthesized according to the expected vector-insert junctional sequence is used as aprobe in colony hybridization screening. Using this strategy, only recombinant clones which fulfill theabove criteria will show positive signals in hybridization.
- 中文關鍵字: insert orientation; oligonucleotide probe; hybridization screening.
- 英文關鍵字: --