The Standardization and Quantification of Nuclear Factor Kappa B p65 by Real-Time Quantitative Polymerase Chain Reaction DOI Open Access
Sivasubramaniyan Gnanaskandan,

Urmila Karunakaran,

Padma Srikanth

et al.

Cureus, Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 29, 2024

The accurate quantification of nuclear factor Kappa B p65 (NF-κB p65) is critical for understanding inflammatory mechanisms, especially in HIV-1 infected individuals, where NF-κB contributes to chronic immune activation. Conventional methods such as enzyme-linked immunosorbent assay (ELISA) and western blotting are limited terms sensitivity reproducibility. This study aimed devise a standardized real-time quantitative polymerase chain reaction (RT-qPCR) using specifically designed primers probe. human sequence was obtained from the National Center Biotechnology Information (NCBI) database, specific probes were Primer3 v4.1.0 software, optimizing melting temperature guanine-cytosine (GC) content ensure stability. Standards synthesized ImmuGenix cloning kit (ImmuGenix Biosciences Pvt. Ltd., Chennai, India) at an initial concentration 2.8 × 10

Language: Английский

The Standardization and Quantification of Nuclear Factor Kappa B p65 by Real-Time Quantitative Polymerase Chain Reaction DOI Open Access
Sivasubramaniyan Gnanaskandan,

Urmila Karunakaran,

Padma Srikanth

et al.

Cureus, Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 29, 2024

The accurate quantification of nuclear factor Kappa B p65 (NF-κB p65) is critical for understanding inflammatory mechanisms, especially in HIV-1 infected individuals, where NF-κB contributes to chronic immune activation. Conventional methods such as enzyme-linked immunosorbent assay (ELISA) and western blotting are limited terms sensitivity reproducibility. This study aimed devise a standardized real-time quantitative polymerase chain reaction (RT-qPCR) using specifically designed primers probe. human sequence was obtained from the National Center Biotechnology Information (NCBI) database, specific probes were Primer3 v4.1.0 software, optimizing melting temperature guanine-cytosine (GC) content ensure stability. Standards synthesized ImmuGenix cloning kit (ImmuGenix Biosciences Pvt. Ltd., Chennai, India) at an initial concentration 2.8 × 10

Language: Английский

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