APPLICATIONS OF MAGNETIC BEADS WITH COVALENTLY ATTACHED OLIGONUCLEOTIDES IN HYBRIDIZATION - ISOLATION AND DETECTION OF SPECIFIC MEASLES-VIRUS MESSENGER-RNA FROM A CRUDE CELL LYSATE

被引：38

作者：

ALBRETSEN, C ^{[1
]}

KALLAND, KH ^{[1
]}

HAUKANES, BI ^{[1
]}

HAVARSTEIN, LS ^{[1
]}

KLEPPE, K ^{[1
]}

机构：

[1] UNIV BERGEN,CTR BIOTECHNOL,N-5014 BERGEN,NORWAY

来源：

ANALYTICAL BIOCHEMISTRY | 1990年 / 189卷 / 01期

基金：

中国国家自然科学基金;

关键词：

D O I：

10.1016/0003-2697(90)90041-7

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A novel, rapid, one-step isolation procedure utilizing oligonucleotides end-attached to magnetic beads (Dynabeads) has been developed. The beads and their captured target nucleic acids were separated after 2 h from the hybridization solution using an external magnet. This procedure was used to isolate measles virus hemagglutinin (H) mRNA from infected cells dissolved in 5 m guanidine thiocyanate. The yield of the specific, undegraded H-mRNA was found to be near the estimated total amount of H-mRNA present in the cells. The magnetic beads were also used for detection of measles virus H-mRNA in a crude cell lysate by sandwich hybridization. The experimental detection limit was 630 amol H-mRNA. © 1990.

引用

页码：40 / 50

页数：11

共 33 条

[1] OPTIMAL CONDITIONS FOR HYBRIDIZATION WITH OLIGONUCLEOTIDES - A STUDY WITH MYC-ONCOGENE DNA PROBES [J].