Sybr green based pcr master mix

Manufactured by Bio-Rad

Sourced in China

SYBR Green-based PCR Master Mix is a ready-to-use solution containing all the necessary components for real-time PCR amplification, including DNA polymerase, dNTPs, and SYBR Green I dye. It is designed to simplify and streamline the setup of real-time PCR reactions.

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Lab products found in correlation

Bulge looptm mirna qpcr primer set, by RiboBio (1 mentions) Cfx96 real time pcr system, by Bio-Rad (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions)

2 protocols using sybr green based pcr master mix

miRNA and mRNA Quantification by qPCR

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For miRNA quantification, the Bulge-LoopTM miRNA qPCR Primer Set (RiboBio, Guangzhou, China) was used to detect miRNA expression with a SYBR Green-based PCR Master Mix (Bio-rad) and a Bio-rad CFX96 Real-Time PCR System. For
marker gene quantification, a 1/10 dilution of cDNA was used as a template with a SYBR Green-based PCR Master Mix (Bio-rad) on a Bio-rad CFX96 Real-Time PCR System. The relative expression levels of miRNAs and mRNAs were
calculated using the 2^–ΔΔCt method and either U6 or glyceraldehyde-3-phosphate dehydrogenase (Gapdh) as internal controls, respectively, using a previously reported protocol [23 (link)].

WU F., TAO L., GAO S., REN L., WANG Z., WANG S., TIAN J, & AN L. (2017). miR-6539 is a novel mediator of somatic cell reprogramming that represses the translation of Dnmt3b. The Journal of Reproduction and Development, 63(4), 415-423.

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Multimodal Experimental Protocols for Cellular Analysis

Cited 1 time

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For western blot assays, cells were harvested in cold PBS solution, and protein lysates were extracted using RIPA buffer as described (11 (link), 12 (link)). After protein assay, equal amounts of proteins from each treatment were subjected to western blot assay with the antibodies indicated in the figure.
For real-time quantitative RT-PCR (qPCR) assays, cells were harvested in cold PBS solution, and total RNAs were extracted using the TRIzol™ reagent (Invitrogen). After the cDNA synthesis, the qPCR reaction was conducted with an SYBR Green-based PCR master mix (Bio-Rad) described in our publications (13 (link), 14 (link)). The house-keeping gene GAPDH was used as an internal control for data normalization.
For luciferase assay, cells were seeded in a 6-well plate overnight and then transfected with an empty reporter construct pGL3 or the human NR0B2 promoter-driven luciferase construct (hSHP-LUC) as described in our publication (14 (link)). After treatment with kinase inhibitors or GW4064 overnight, cells were harvested for luciferase measurement, and the final readings were normalized with the relative levels of total proteins, as described in our publication (15 (link)).

Zhu R., Tu Y., Chang J., Xu H., Li J.C., Liu W., Do A.D., Zhang Y., Wang J, & Li B. (2021). The Orphan Nuclear Receptor Gene NR0B2 Is a Favorite Prognosis Factor Modulated by Multiple Cellular Signal Pathways in Human Liver Cancers. Frontiers in Oncology, 11, 691199.

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