摘要
以M13mp19为模板,寡核苷酸“1224”为引物,进行半随机单引物PCR扩增;分析其扩增产物的限制性内切酶酶切图谱,推定它们分别在M13mp19序列中的确切位置;证实引物“1224”的3端与模板M13mp19负链的互补结合,至少需有连续的4个核苷酸,才能产生单引物PCR扩增的模板分子,进行有效的PCR扩增,并由此决定了扩增产物中各DNA片段的大小及其在模板DNA序列中的特定位置.
The template M,13mp19 was amplified by the semi-random single-primer PCR with the oligonucleotide primer '1224', The DNA fragments in the amplification products were purified and used as the templates. The latter was then amplified by the same single-primer PCR under the normal condition. The exact regions where these products were located and the starting sites of the specific PCR amplification were determined. It was shown that more than 4 nucleotides at the 3,termination of the primer'1224' integrating complementarily with the template DNA were needed for the production of the template molecules of single-primer PCR and the effective PCR amplification. Moreover, the sites of integration determined the lengths of DNA fragments in the amplification products and their specific locations in the template DNA.
出处
《复旦学报(自然科学版)》
CAS
CSCD
北大核心
1994年第2期121-126,共6页
Journal of Fudan University:Natural Science
关键词
扩增
聚合酶
遗传工程
单引物
PCR
primer, amplification
polymcrase
restriction endonucleases,polymerasc chain reaction
single-primer PCR amplification