Quantitative real-time PCR for rapid and accurate titration of recombinant baculovirus particles

被引：60

作者：

Hitchman, Richard B. ^{[1
]}

Siaterli, Evangelia A. ^{[1
]}

Nixon, Clare P. ^{[1
]}

King, Linda A. ^{[1
]}

机构：

[1] Oxford Brookes Univ, Sch Biol & Mol Sci, Oxford OX3 0BP, England

来源：

BIOTECHNOLOGY AND BIOENGINEERING | 2007年 / 96卷 / 04期

基金：

英国生物技术与生命科学研究理事会;

关键词：

quantitative PCR (QPCR); baculovirus; titer; high throughput;

D O I：

10.1002/bit.21177

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

We describe the use of quantitative PCR (QPCR) to titer recombinant baculoviruses. Custom primers and probe were designed to gp64 and used to calculate a standard curve of QPCR derived titers from dilutions of a previously titrated baculovirus stock. Each dilution was titrated by both plaque assay and QPCR; producing a consistent and reproducible inverse relationship between C-T and plaque forming units per milliliter. No significant difference was observed between titers produced by QPCR and plaque assay for 12 recombinant viruses, confirming the validity of this technique as a rapid and accurate method of baculovirus titration. (c) 2006 Wiley Periodicals, Inc.

引用

页码：810 / 814

页数：5

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