IMAGE-ANALYSIS IN QUANTITATIVE PCR - AN APPLICATION FOR THE MEASUREMENT OF C-ERBB-2 ONCOGENE AMPLIFICATION IN DNA FROM HUMAN TUMORS

被引：21

作者：

ORLANDO, C ^{[1
]}

SESTINI, R ^{[1
]}

ZENTILIN, L ^{[1
]}

GELMINI, S ^{[1
]}

PINZANI, P ^{[1
]}

GIACCA, M ^{[1
]}

PAZZAGLI, M ^{[1
]}

机构：

[1] INT CTR GENET ENGN & BIOTECHNOL,TRIESTE,ITALY

来源：

JOURNAL OF BIOLUMINESCENCE AND CHEMILUMINESCENCE | 1994年 / 9卷 / 03期

关键词：

PCR; COMPETITIVE PCR; DNA; IMAGE ANALYSIS; ONCOGENES; TUMOR;

D O I：

10.1002/bio.1170090317

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We present an application of image analysis for the direct quantification of PCR products after gel electrophoresis and ethidium bromide staining of DNA. This procedure has been applied to the development of an assay based on competitive PCR for the measurement of the degree of amplification of c-erbB-2 oncogene in DNA from human tumours. In this method two DNA species (genomic and competitor) compete for PCR amplification. Since results are calculated from the final competitor/genomic ratio any variable affecting the rate of PCR amplification has no effect on the accuracy of the ratio measurement. Results are reported which show that even large variations in the experimental conditions (number of PCR cycles, sample volumes and extracted DNA quality) did not interfere with the precision of the measurement of the competitor/genomic ratio.

引用

页码：223 / 228

页数：6

共 16 条

[1] MOLECULAR PROFILE OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 INFECTION IN SYMPTOMLESS PATIENTS AND IN PATIENTS WITH AIDS [J].