Sybr green system

Manufactured by Takara Bio

Sourced in China, Japan, United States, Canada

The SYBR Green system is a real-time PCR detection method that utilizes the SYBR Green dye to quantify the amount of DNA in a sample. The dye binds to double-stranded DNA, and the fluorescence intensity increases as the DNA amplifies during the PCR process, allowing for real-time monitoring of the reaction.

Automatically generated - may contain errors

Lab products found in correlation

Trizol reagent, by Thermo Fisher Scientific (7 mentions) Trizol, by Thermo Fisher Scientific (4 mentions) Trizol reagent, by Takara Bio (4 mentions) Rnaiso plus, by Takara Bio (4 mentions) Primescript rt reagent kit, by Takara Bio (4 mentions) Rneasy mini kit, by Qiagen (4 mentions) Nanodrop 1000, by Thermo Fisher Scientific (4 mentions) Primescript rt master mix, by Takara Bio (3 mentions) Trizolr reagent, by Thermo Fisher Scientific (2 mentions) Abi 7500, by Thermo Fisher Scientific (2 mentions) Reverse transcription reagent, by Takara Bio (2 mentions) Lightcycler 480, by Takara Bio (2 mentions) Beacon designer 7, by Premier Biosoft (2 mentions) Steponeplus rt pcr system, by Thermo Fisher Scientific (2 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (2 mentions) Abi prism 7900 sequence detection system, by Thermo Fisher Scientific (2 mentions) Cfx96 real time pcr system, by Bio-Rad (1 mentions) Dneasy kit, by Qiagen (1 mentions) Trizol, by Takara Bio (1 mentions) Lightcycler 480, by Roche (1 mentions)

Close spelling variants of this product

SYBR Green real-time PCR system SYBR Green qPCR Master Mix reagent system SYBR Green PCR Master Mix system SYBR Green qPCR system SYBR Green detection reagent system SYBR Green Premix Ex Taq™ II quantitative PCR system SYBR green detection system (Bio SYBR Green Master Mix SYBR Green reaction system SYBR green detection RT-PCR system SYBR green detection system

40 protocols using sybr green system

Quantitative PCR Analysis of TERT and Telomere Length in NPCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was isolated from NPCs using TRIzol^® reagent (Invitrogen; Thermo Fisher Scientific, Inc.), and 1 µg of total RNA was used to synthesize cDNA. Quantitative PCR was performed using the SYBR^® Green system (Takara Bio, Inc.). Amplification of cDNA samples was carried out on the CFX96 real-time PCR system (Bio-Rad Laboratories, Inc.). The reaction steps were as follows: 95˚C for 3 min, 60˚C for 45 sec and for 39 cycles. At-last melt curve analysis: 65˚C Then 95˚C and 5˚C for 5 sec each. The relative quantification of the target gene was normalized to β-actin, and target gene was compared with the control sample using the 2^-ΔΔCq method (24 (link)). The primers for TERT and β-actin are listed in Table I.
DNA was extracted using the DNeasy kit from Qiagen GmbH. After the DNA was extracted from the NPCs in six-well plates at a seeding density of 2x10⁵ cells/ml following the manufacturer's protocols, then the same protocol and quantification of qPCR data were carried out as aforementioned. The primers of Telomere and 36B4 are listed in Table I.

Hong H., Xiao J., Guo Q., Du J., Jiang Z., Lu S., Zhang H., Zhang X, & Wang X. (2021). Cycloastragenol and Astragaloside IV activate telomerase and protect nucleus pulposus cells against high glucose-induced senescence and apoptosis. Experimental and Therapeutic Medicine, 22(5), 1326.

+ Open protocol

+ Expand

Quantification of Inflammatory Cytokine Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA from the synovial tissue of CIA mice or culture cells was extracted with TRIzol Rreagent (Invitrogen) and reverse transcripted into cDNA. Transcription levels of IL-4, IFN-γ, IL-17A, ROR-γt, T-bet and GATA3 genes were analysed by real-time quantitative PCR using an ABI 7500 (Applied Biosystems) and SYBR green system (TaKaRa), according to the manufacturer’s protocol. The primer sequences are summarised in Supplemental Table 2. The relative gene expressions were normalised to the level of β-action and quantified by the 2^−ΔΔCT method. All reactions were performed in duplicate for each sample.

Liu D., Cao T., Wang N., Liu C., Ma N., Tu R, & Min X. (2016). IL-25 attenuates rheumatoid arthritis through suppression of Th17 immune responses in an IL-13-dependent manner. Scientific Reports, 6, 36002.

+ Open protocol

+ Expand

Quantitative RT-PCR Gene Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was isolated with Trizol (Takara Bio Inc., Beijing, China) following the manufacturer’s protocol. Reverse transcription reagents (Takara Bio Inc.) were applied to reverse transcribe RNA. qRT-PCR was carried out to quantify the target genes’ expression with SYBR Green system (Takara Bio Inc.).

Wang Y., Zhou J.S., Xu X.C., Li Z.Y., Chen H.P., Ying S.M., Li W., Shen H.H, & Chen Z.H. (2018). Endoplasmic reticulum chaperone GRP78 mediates cigarette smoke-induced necroptosis and injury in bronchial epithelium. International Journal of Chronic Obstructive Pulmonary Disease, 13, 571-581.

+ Open protocol

+ Expand

Quantification of A20 Gene Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was extracted by Trizol reagent (Invitrogen) according to the manufacturer's instructions. The first-strand cDNA was synthesized using oligo(dT) primers and the RevertAid™ M-MuLV transcription enzyme (Fermentas, Pittsburgh, PA, USA). Samples were subjected to real-time RT-PCR analysis on a Lightcycler480 (Roche Diagnostics, Mannheim, Germany) with SYBR Green system (Takara, Biotechnology, Dalian, China) using the following parameters: 2 min. at 50°C followed by 10 min. at 95°C and 40 cycles of 15 sec. at 95°C and 1 min. at 60°C. Gene expression levels were normalized to that of housekeeping gene GAPDH. Primers were as follows: A20 Sense: 5′-CTAAGCCAACGAGTAGGTTCTGTG-3′, Antisense: 5′-CCATACA TCTGCTTGAACTGGTAG-3′; GAPDH Sense: 5′-CTCTGGAAAGCTGTGG CGTGATG-3′, Antisense: 5′-ATGCCAGTGAGCTTCCCGTTCAG-3′.

Gui J., Chen R., Xu W, & Xiong S. (2015). Remission of CVB3-induced myocarditis with Astragaloside IV treatment requires A20 (TNFAIP3) up-regulation. Journal of Cellular and Molecular Medicine, 19(4), 850-864.

+ Open protocol

+ Expand

Quantifying Gene Expression by Real-Time PCR

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA were extracted from collected cells by Trizol (Invitrogen) and reverse‐transcribed (RT) using a cDNA synthesis kit (TaKaRa, China) according to the manufacturer's instructions. The expression of the genes encoding AIM2, ASC, caspase‐1 and IL‐1β was quantified by real‐time PCR using 7900HT qPCR system thermal cycler (Applied Biosystems) and SYBR Green system (TaKaRa) normalized by GAPDH expression following the manufacturer's protocol. Primers were used as following in Table 1.

Chai D., Qiu D., Zhang Z., Yuchen Shi S., Wang G., Fang L., Li H., Li H., Tian H, & Zheng J. (2020). Absent in melanoma 2 enhances anti‐tumour effects of CAIX promotor controlled conditionally replicative adenovirus in renal cancer. Journal of Cellular and Molecular Medicine, 24(18), 10744-10755.

+ Open protocol

+ Expand

Analyzing gene expression in mouse lung

Check if the same lab product or an alternative is used in the 5 most similar protocols

Extraction of total RNA in the lung was done with TRIzol reagent (TRIzol reagent, TAKARA Bio Inc.) in accordance with the specification, then we conducted reverse transcription polymerase chain reaction on the complementary DNA samples using the SYBR Green system (TAKARA Bio Inc.). Mouse TATA-binding protein (TBP) was used as an endogenous control for gene expression normalization. The fold changes were calculated using the ΔΔCt method of relative comparison. The sequence for the primer sets used is as follows. The primers used were P2X4R sense 5’-ATCGTCACCGTGAACCAGAC-3’ and P2X4R antisense 5’-GCGTCTGAATCGCAAATGCT-3’, GATA-3 sense 5’-CTTATCAAGCCCAAGCGAAG-3’ and GATA-3 antisense 5’-CCCATTAGCGTTCCTCCTC-3’, T-bet sense 5’-TCAACCAGCACCAGACAGAG-3’ and T-bet antisense 5’-AACATCCTGTAAT GGCTTGTG-3’, TBP sense 5’-GTGGATCGAGTCCGGTA GC-3’, TBP antisense 5’-AAT AGTGATGCTGGGCACTG-3’. Cycle and threshold were obtained in accordance with the specification of the manufacturer.

Hu B., Feng X., Wang L., Song Y, & Ni X. (2018). 5-BDBD ameliorates an OVA-induced allergic asthma by the reduction of Th2 cytokines production. Iranian Journal of Basic Medical Sciences, 21(4), 364-369.

+ Open protocol

+ Expand

Quantifying miRNA-372-3p Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was isolated by an RNA extraction kit (Solarbio, China) and reverse transcribed into cDNA by PrimeScript RT Master Mix (Takara Bio). qRT-PCR was performed with a SYBR Green system (Takara) according to the following parameters: 95°C for 10 min, followed by 40 cycles of 95°C for 10 s, and 60°C for 50 s. For miRNA expression, U6 were used as internal reference controls for miRNA expression, respectively, and relative expression was calculated using the 2^−ΔΔCt method and was used to calculate the relative expression levels of miR-372-3p. Primers: miR-372-3p, forward 5′-TTT CAC GAC GCT GTA AAC TCG CA-3′, reverse 5′ -GTG CAG GGT CCG AGG T-3′; U6, forward 5′-GCT TCG GCA GCA CAT ATA CTA A-3′, reverse 5′-AAC GCT TCA CGA ATT TGC GT-3′.

Han Z., Zhao D., Han M., Zhang R, & Hao Y. (2022). Knockdown of miR-372-3p Inhibits the Development of Diabetic Cardiomyopathy by Accelerating Angiogenesis via Activating the PI3K/AKT/mTOR/HIF-1α Signaling Pathway and Suppressing Oxidative Stress. Oxidative Medicine and Cellular Longevity, 2022, 4342755.

+ Open protocol

+ Expand

Quantitative Gene Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was extracted using TRIzol reagent (Takara Bio, Inc., Shiga, Japan; #3732) according to the manufacturer’s instructions. RNA concentrations were equalized and converted to cDNA using a kit (Takara; #RR037). Gene expression was measured by qPCR (Applied Biosystems, Thermo Fisher Scientific, Carlsbad, CA, USA) using the SYBR Green system (Takara Bio, Inc.). Expression was normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) or β-Actin. Primer information is provided in Supplementary Table S1. All gene expression procedures, including the design of primers, quantitation methods, and validation of PCR environment, were conducted according to previous studies (Johnson et al., 2014 (link)).

Yu S., Jiang J., Li Q., Liu X., Wang Z., Yang L, & Ding L. (2022). Schisantherin A alleviates non-alcoholic fatty liver disease by restoring intestinal barrier function. Frontiers in Cellular and Infection Microbiology, 12, 855008.

+ Open protocol

+ Expand

Quantitative RT-PCR Analysis of HBV RNA and HNF4α

Check if the same lab product or an alternative is used in the 5 most similar protocols

qRT-PCR was performed as described previously [28 (link), 30 (link)]. Briefly, the cells or liver tissues were harvested, and total RNA was isolated with RNAiso Plus (Takara, Dalian, China). cDNA was synthesized from 0.5 μg of RNA using the PrimeScript™ RT Reagent Kit with gDNA Eraser (Perfect Real Time) (Takara). Prior to reverse transcription, gDNA Eraser was used to remove contaminating DNA according to the manufacturer’s instructions. The qRT-PCR analysis of cDNA targets was performed using a Lightcycler 480 and a SYBR Green system (Takara). The primers used in this study are as follows: HBV pgRNA, forward, 5′- TGTTCAAGCCTCCAAGCT-3′ and reverse, 5′-GGAAAGAAGTCAGAAGGCAA-3′; HBV RNA, forward, 5′-GCACTTCGCTTCACCTCTGC-3′ and reverse, 5′-CTCAAGGTCGGTCGTTGACA-3′; HNF4α, forward, 5′-TGTCCCGACAGATCACCTC-3′ and reverse, 5′-CACTCAACGAGAACCAGCAG-3′; GAPDH, forward, 5′-CGGGTGGGAATGTTGAGG-3′ and reverse, 5′-TGGCGGGAGATGTGGGTAC-3′. The levels of pgRNA, HBV RNA or HNF4α were calculated following normalization to GAPDH levels by the comparative ΔΔ threshold cycle method. The specificity of the amplification reactions was confirmed by melt curve analysis. The results are representative of three independent experiments.

Wang Z.Y., Li Y.Q., Guo Z.W., Zhou X.H., Lu M.D., Xue T.C, & Gao B. (2019). ERK1/2-HNF4α axis is involved in epigallocatechin-3-gallate inhibition of HBV replication. Acta Pharmacologica Sinica, 41(2), 278-285.

+ Open protocol

+ Expand

Evaluating Transcriptional Levels of EDIL3, XIST, and miR-137

Check if the same lab product or an alternative is used in the 5 most similar protocols

The transcription levels of EDIL3, XIST and miR-137 were evaluated by RT-qPCR assay. A commercial RNA extraction kit obtained from TaKaRa Bio Co. Ltd. was used to extract RNA from indicated tissues and cell lines. Next, the total RNA was produced into cDNA using a Prime Script RT Master Mix (Takara, China). Thirdly, The RT-qPCR assay was carried out using a SYBR-Green system (Takara), in agreement with the kit’s instructions. Finally, the 2^−ΔΔCt method was performed to obtain the transcriptional levels of EDIL3, XIST and miR-137. The glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the internal reference gene for EDIL3 and XIST, and U6 was used as the internal reference gene for miR-137. The primer sequences of EDIL3, XIST, miR-137, U6 and GAPDH used in this study were showed in Table 2.

Zhang L., Peng K.W., Wang B., Yang X.F, & Zhang Z.M. (2020). EDIL3 regulates gastric cancer cell migration, invasion and epithelial-mesenchymal transition via TGF-β1/XIST/miR-137 feedback loop. Translational Cancer Research, 9(10), 6313-6330.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!