Quantitative Determination of Lentiviral Vector Particle Numbers by Real-Time PCR

Autor:	Michaela Scherr, U Blömer, M Grez, Arnold Ganser, Karin Battmer
Rok vydání:	2001
Předmět:	Biology Virology General Biochemistry Genetics and Molecular Biology Quantitative determination Viral vector Running time law.invention Real-time polymerase chain reaction Capsid law Complementary DNA Particle Biological system Polymerase chain reaction Biotechnology
Zdroj:	BioTechniques. 31:520-526
ISSN:	1940-9818 0736-6205
Popis:	Here, we describe a quantitative, DNAbased, real-time PCR approach to determine the number of lentivirus particles that are present in vector preparations. In this approach, the minus strong-stop cDNA fragment that is present in viral capsids serves as template for PCR. Using this technology, we found that only 0.1%–1% of the virus particles that are present in vector preparations are infectious. The approach described here is rapid, reliable, and simple in concept and can be used to estimate both vector particles in supernatants and the number of infectious particles. Also, this approach can easily be adapted to a highthroughput system by using 96-well plates and a 2-h running time.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::97054542290565c60dc45c5a61c7dfa4 https://doi.org/10.2144/01313st05 Zobrazit plný text záznamu