Real-time quantitative PCR for the design of lentiviral vector analytical assays
نویسندگان
چکیده
منابع مشابه
Quantitative determination of lentiviral vector particle numbers by real-time PCR.
Here, we describe a quantitative, DNA-based, 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 infectio...
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The ability to multiplex PCR by probe color and melting temperature (T(m)) greatly expands the power of real-time analysis. Simple hybridization probes with only a single fluorescent dye can be used for quantification and allele typing. Different probes are labeled with dyes that have unique emission spectra. Spectral data are collected with discrete optics or dispersed onto an array for detect...
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lentiviral vectors (lvs) are useful vehicle for genetransfer to dividing and non-dividing cells and genetic manipulations. however, the use of lentiviruses in studies requires an accurate titration technique.quantitative real-time pcr (qpcr) is a sensitive technique for the indication and quantitation of retrovirals particles. in this study, we used the qpcr for lentiviral vector titeration. th...
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Unlike classical end-point analysis PCR, real-time PCR provides the data required for quantification of the target nucleic acid. The results can be expressed in absolute terms by reference to external quantified standards or in relative terms compared to another target sequence present within the sample. Absolute quantification requires that the efficiency of the amplification reaction is the s...
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We have developed a novel "real time" quantitative PCR method. The method measures PCR product accumulation through a dual-labeled fluorogenic probe (i.e., TaqMan Probe). This method provides very accurate and reproducible quantitation of gene copies. Unlike other quantitative PCR methods, real-time PCR does not require post-PCR sample handling, preventing potential PCR product carry-over conta...
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ژورنال
عنوان ژورنال: Gene Therapy
سال: 2005
ISSN: 0969-7128,1476-5462
DOI: 10.1038/sj.gt.3302614