RNA was extracted from brain, eye and spleen tissue samples using RNeasy Plus Mini Kit (QIAgen), following the manufacturer’s recommendations. Viral RNA was quantified using One Step Taqman RT-qPCR (Thermo Fisher Scientific) on a 7500 Real-Time PCR System (Applied Biosystems) with primers and probe designed by Lanciotti and collaborators44 (link). ZIKV copies were calculated by interpolation onto an internal standard curve formed of eight 10-fold serial dilutions of a synthetic ZIKV RNA based on the Asian lineage. The ZIKV quantification was displayed in ZIKV RNA copies per gram of tissue.
Garcez P.P., Stolp H.B., Sravanam S., Christoff R.R., Ferreira J.C., Dias A.A., Pezzuto P., Higa L.M., Barbeito-Andrés J., Ferreira R.O., Andrade C.B., Siqueira M., Santos T.M., Drumond J., Hoerder-Suabedissen A., de Lima C.V., Tovar-Moll F., Lopes R.T., Fragel-Madeira L., Lent R., Ortiga-Carvalho T.M., Stipursky J., Bellio M., Tanuri A, & Molnár Z. (2018). Zika virus impairs the development of blood vessels in a mouse model of congenital infection. Scientific Reports, 8, 12774.
Corresponding Organization : Universidade Federal do Rio de Janeiro
Other organizations :
University of Oxford, Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro, D’Or Institute for Research and Education, Universidade Federal Fluminense
RNA extraction using RNeasy Plus Mini Kit (QIAgen)
Viral RNA quantification using One Step Taqman RT-qPCR (Thermo Fisher Scientific) on a 7500 Real-Time PCR System (Applied Biosystems)
Primers and probe designed by Lanciotti and collaborators
Internal standard curve formed of eight 10-fold serial dilutions of a synthetic ZIKV RNA based on the Asian lineage
controls
Positive control: Synthetic ZIKV RNA based on the Asian lineage
Negative control: Not specified
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to
get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
