Quantification of M13 and T7 bacteriophages by TaqMan and SYBR green qPCR.

ABSTRACT: TaqMan and SYBR Green quantitative PCR (qPCR) methods were developed as DNA-based approaches to reproducibly enumerate M13 and T7 phages from phage display selection experiments individually and simultaneously. The genome copies of M13 and T7 phages were quantified by TaqMan or SYBR Green qPCR referenced against M13 and T7 DNA standard curves of known concentrations. TaqMan qPCR was capable of quantifying M13 and T7 phage DNA simultaneously with a detection range of 2.75*10¹-2.75*10⁸genome copies(gc)/μL and 2.66*10¹-2.66*10⁸ genome copies(gc)/μL respectively. TaqMan qPCR demonstrated an efficient amplification efficiency (E_s) of 0.97 and 0.90 for M13 and T7 phage DNA, respectively. SYBR Green qPCR was ten-fold more sensitive than TaqMa

SUBMITTER: Peng X 

PROVIDER: S-EPMC5745055 | biostudies-literature | 2018 Feb

REPOSITORIES: biostudies-literature