耐热FD DNA多聚酶在PCR中的应用条件

被引：4

作者：

毛裕民

杨树青

唐榕

刘坚

冯霄

马匆匆

机构：

[1] 复旦大学遗传学研究所

来源：

复旦学报(自然科学版) | 1991年 / 02期

关键词：

链式反应,DNA扩增,聚合酶; 多聚酶链反应;

D O I：

10.15943/j.cnki.fdxb-jns.1991.02.010

中图分类号：

学科分类号：

摘要：

探索了用于多聚酶链式反应（PCR）的FD DNA多聚酶的最适反应条件在100μl的反应液中含有25 mmol/L的Tris-HCl（pH8.0）,25 mmol/L的（NH4）2SO4,1.5 mmol/L的MgCl2,200 μg/ml的明胶,dNTP各200 μmol/L,模板量为3.3×10～（-2）Pg,引物量各为290 nmol/L,1～2 U的耐热FD DNA多聚酶,30次循环反应的系统为通用PCR最适反应系统。应用这一反应系统,混迹于大量DNA分子中的特定DNA片段可借助于琼脂糖凝胶电泳专一性地、快速而有效地检出。

引用

页码：186 / 193

页数：8

共 11 条

[1]

Direct Cloning and Sequence Analysis of Enzymztically Amplified Genomic Sequences. Scharf S J,et al. Science . 1986

[2]

DNA Typing From Single Hairs. Higuchi R,et al. Nature . 1988

[3]

Enzymatic Amplification of β-globin Genomic Sequences and Restriction Site Analysis for Dignosis of Sickle cell Anemia. Saiki R K,et al. Science . 1985

[4]

Enzymatic Synthesis of Deoxyribonucleic Acid--VII.Synthesis of a Polymer of Deoxyadenylate and Deoxythymidylate. Schachman N K,et al. Journal of Biological Chemistry . 1960

[5]

Characterization or β-Thalassaemia Mutations Using Direct Genomic Sequencing of Amplified Single Copy DNA. Wong C,et al. Nature . 1987

[6]

Identification of Human Immunodeficiency Virus Sequences by Using in Vitro Enzymatic Amplification and Oligomer Cleavage Detection. Kwok S,et al. Journal of Virology . 1987

[7]

Primer-directed Emzymatic Amplification of DNA With a Thermostable DNA Polymerase. Saiki R K,et al. Science . 1988

[8]

Delection of Minimal Residual cells Carrying the t(14: 18) by DNA Sequence Amplification. Lee M S,et al. Science . 1987

[9]

DNA Diagnostics--Molecular Techniques and Automation. Landegren U,et al. Science . 1988

[10]

Characterization of c-ki-ras Oncogene Alleles by Direct Sequencing of Enzymatically Amplified DNA From Carcinogen-induced Tumors. Mcmanon G,et al. Proceedings of the National Academy of Sciences of the United States of America . 1987

← 1 2 →