Regular ArticleA Quantitative Method of Determining Initial Amounts of DNA by Polymerase Chain Reaction Cycle Titration Using Digital Imaging and a Novel DNA Stain
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Advances in the diagnosis of key gastrointestinal nematode infections of livestock, with an emphasis on small ruminants
2013, Biotechnology AdvancesCitation Excerpt :The principle of the original method was to incorporate a specific, intercalating dye (e.g., ethidium bromide) into the PCR to measure the change in fluorescence after each cycle using a digital camera and a fluorometer coupled to the reaction tube (Higuchi et al., 1993). The technique has been modified to include other (non-carcinogenic) dyes, such as SYBR Green I (Becker et al., 1996), LCGreen (Wittwer et al., 2003), SYTO9 (Monis et al., 2005a) and EvaGreen (Wang et al., 2006). Real-time PCR assays using such dyes enable the relative or absolute quantitation of amplicons by allowing the identification of the cycle (Ct) at which the amplification commences.
Unique gangliosides synthesized in vitro by sialyltransferases from marine bacteria and their characterization: Ganglioside synthesis by bacterial sialyltransferases
2013, Journal of Lipid ResearchCitation Excerpt :As a final step, they were visualized with peroxidase substrate solution (immunostain kit, Konica, Tokyo). Sialylated products were analyzed by in situ polyvinylidene difluoride (PVDF) membrane (ATTO, Tokyo, Japan) transfer followed by secondary ion mass spectrometry (SI-MS), as described previously (26, 27). GSLs developed by TLC were transferred to a PVDF membrane by TLC blotting, after which GSL bands on the membrane were developed with primurin spray (28), excised, and placed on a mass spectrometer probe tip, with a few microliters of triethanolamine added as the matrix.
Next-Generation Molecular-Diagnostic Tools for Gastrointestinal Nematodes of Livestock, with an Emphasis on Small Ruminants. A Turning Point?
2013, Advances in ParasitologyCitation Excerpt :The principle of the original method was to incorporate a specific, intercalating dye (e.g. ethidium bromide) into the PCR to measure the change in fluorescence after each cycle using a digital camera and a fluorometer coupled to the reaction tube (Higuchi et al., 1993). The technique has been modified to include other (non-carcinogenic) dyes, such as SYBR Green I (Becker et al., 1996), LCGreen (Wittwer et al., 2003), SYTO9 (Monis et al., 2005a) and EvaGreen (Wang et al., 2006). RT-PCR assays using such dyes enable the relative or absolute quantitation of amplicons by allowing the identification of the cycle (Ct) at which the amplification commences.
Toward routine, DNA-based detection methods for marine pests
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