SYBR Green qPCR protocol Version 1
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SYBR Green qPCR Protocol.docx
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Created: 22nd Nov 2023 at 22:55
Last updated: 3rd Jan 2024 at 17:38
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Version 1 (earliest) Created 22nd Nov 2023 at 22:55 by John Cornelius
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https://orcid.org/0009-0003-6381-9603
Public web page: Not specified
Submitter: John Cornelius
Studies: Viral Protein Accumulation of Zika Virus Variants Links with Regulation ...
Assays: Bioinformatics analyses of NanoString data, Deep sequencing of ZIKV stocks and sequence alignment, FFU assay for determining the titers of ZIKV stocks, FFU assay for measuring viral spread, Flow cytometry, Immunoblot analysis, Immunofluorescence analysis, Plaque assay, Protein lysate quantification, Quantification of extracellular and intracellular RNA, Quantification of extracellular and intracellular virions, Sample - Metadata, Transcript analysis by NanoString, Transcript analysis by SYBR green qRT-PCR, Viral copy number quantification by TaqMan qRT-PCR, ZIKV attachment and entry assay, ZIKV infection, ZV-13 antibody purification and quantification
Asian lineage Zika virus (ZIKV) strains emerged globally, causing outbreaks linked with critical clinical disease outcomes unless the virus is effectively restricted by host immunity. We have previously shown that retinoic acid-inducible gene-I (RIG-I) senses ZIKV to trigger innate immunity to direct interferon (IFN) production and antiviral responses that can control ZIKV infection. However, ZIKV proteins have been demonstrated to antagonize IFN. Here, we conducted in vitro analyses to assess ...
Submitter: John Cornelius
Investigation: Zika Virus
Assays: Bioinformatics analyses of NanoString data, Deep sequencing of ZIKV stocks and sequence alignment, FFU assay for determining the titers of ZIKV stocks, FFU assay for measuring viral spread, Flow cytometry, Immunoblot analysis, Immunofluorescence analysis, Plaque assay, Protein lysate quantification, Quantification of extracellular and intracellular RNA, Quantification of extracellular and intracellular virions, Sample - Metadata, Transcript analysis by NanoString, Transcript analysis by SYBR green qRT-PCR, Viral copy number quantification by TaqMan qRT-PCR, ZIKV attachment and entry assay, ZIKV infection, ZV-13 antibody purification and quantification
cDNA was synthesized as described above. For each sample, cDNA was diluted to a final concentration of 2.5 ng/μL for SYBR green qRT-PCR on the QuantStudio 5 real-time PCR system using the SYBR green master mix (Thermo Fisher) and gene-specific primers (Table 4; Qiagen; IDT). Each sample was run in triplicate.
Submitter: John Cornelius
Assay type: Experimental Assay Type
Technology type: qRT-PCR
Investigation: Zika Virus
