The Analogy Between the Effect of Various Dyes for DNA Quantification in QUBIT 4.0
Sowmya Seshadri
Sowmya Seshadri, Department of Chemical Engineering, SRM Institute of Science and Technology, Chennai (Tamil Nadu), India.
Manuscript received on 02 April 2019 | Revised Manuscript received on 18 April 2019 | Manuscript Published on 30 April 2019 | PP: 20-24 | Volume-7 Issue-6S6 April 2019 | Retrieval Number: F10060476S619/2019©BEIESP | DOI: 10.35940/ijrte.F1006.0476S619
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© The Authors. Blue Eyes Intelligence Engineering and Sciences Publication (BEIESP). This is an open access article under the CC-BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)
Abstract: This study has examined the effect of various fluorescent dyes, to check its feasibility for it’s use in a highly sensitive fluorometer. QUBIT 4.0 is a fluorometer that is used to measure the quantity of RNA, DNA or Protein. The QUBIT 4.0 was developed to measure the RNA integrity and quality using its high sensitivity assays. It was also used to measure dilute sample concentrations ranging from 1-20 micro litres; which is why dye selection is crucial. The evolution of dye selection has been discussed in detail, ranging from Ethidium Bromide (commonly used as a fluorescent tag for gel electrophoresis), to the dye chosen here for DNA quantification in QUBIT 4.0, called Pico Green (ultra sensitive dye which is used for ds – DNA quantification in QUBIT 4.0). An alternative dye is also discussed called C-phycocyanine, which ideally has no literature data cited for being used in QUBIT 4.0; but proof of concept points it out to be feasible as it conforms to most of the properties required for the same. Based on this study, one can understand the main features of QUBIT 4.0, the various dyes used in fluorometers and their key properties and also know what criteria to choose while selecting a dye for a particular assay, about Pico Green, the main emphasis laid here, and its alternative; C-phycocyanine.
Keywords: QUBIT 4.0, Assay, Ethidium Bromide, SYBR dyes, Hoescht 33258, PicoGreen, C-phycocyanine, Fluorescence, Jablonski Diagram, Intercalation, Groove Binding, Major Groove, Minor Groove.
Scope of the Article: Bio-Science and Bio-Technology