Sybr green sybr premix ex taq 2

Manufactured by Takara Bio

Sourced in Japan

SYBR Green (SYBR Premix Ex Taq II) is a fluorescent dye used in real-time PCR (qPCR) for the detection and quantification of DNA. It binds to double-stranded DNA and emits a fluorescent signal that can be measured and used to monitor the amplification of target DNA sequences during the PCR process.

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Lab products found in correlation

Nanodrop 2000 spectrophotometer, by Thermo Fisher Scientific (4 mentions) Primescript rt reagent kit, by Takara Bio (3 mentions) Quantstudio 12k flex real time pcr system, by Thermo Fisher Scientific (2 mentions) Trizol reagent, by Thermo Fisher Scientific (2 mentions) Abi prism 7500, by Takara Bio (1 mentions) Tri reagent, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

SYBR Green Premix Ex TaqTM II SYBR Green PCR Premix Ex TaqTM II reagents SYBR Green Premix Ex Taq II SYBR Green Premix Ex Taq II kit SYBR Green PCR Premix Ex Taq II reagents SYBR Green Premix Ex Taq™ II quantitative PCR system SYBR Green Premix Ex Taq II (Tli RNase H Plus, TB SYBR green Premix Ex Taq II SYBR Green Premix Ex TaqTM SYBR Green Premix Ex Taq

4 protocols using sybr green sybr premix ex taq 2

Evaluating Visfatin and Lipocalin-2 Expression in PTB

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In total, 5 mL anticoagulated peripheral blood was extracted from each PTB patient and normal control, and PBMCs were isolated from peripheral blood using Ficoll‐Hypaque density gradient centrifugation. Then, total RNA in PBMCs was extracted by TRIzol reagent, and the RNA concentration was measured using the NanoDrop 2000 spectrophotometer. Finally, total RNA was reverse‐transcribed to cDNA with a PrimeScriptTM RT reagent Kit.
Visfatin and lipocalin‐2 mRNA expression levels in PBMCs were measured using qRT‐PCR with SYBR Green (SYBR Premix Ex Taq II, Takara Bio Inc), and the housekeeping gene β‐actin was used as an internal control to calculate the relative expression levels of these adipokines in the same sample. The relative expression levels of visfatin and lipocalin‐2 were estimated using 2^−△△Ct normalized to β‐actin.²⁰

Li H., Huang Q., Chen S., Zhang G., Shi S., Hua W.a.n.g., Li Y, & Zhang T. (2020). The mRNA expression of visfatin and lipocalin‐2 in peripheral blood mononuclear cells from patients with pulmonary tuberculosis. Journal of Clinical Laboratory Analysis, 34(11), e23476.

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METTL3, METTL14, and WTAP Expression Analysis

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Peripheral blood mononuclear cells (PBMCs) were isolated from 3 mL anticoagulant peripheral blood, and total RNA was extracted using TRIzol Reagent (Invitrogen, Carlsbad, CA, USA). A NanoDrop 2000 spectrophotometer (Thermo Scientific, USA) was used to determine total RNA concentrations. Total RNA was then reverse transcribed into cDNA by the PrimeScript™ RT Reagent Kit (Takara Bio Inc., Japan). In this experiment a 20-μL reverse transcription reaction system with a maximum of 1 μg total RNA was used, and the amount of total RNA needed in the reaction system was determined based on the RNA concentration.
METTL3, METTL14, and WTAP mRNA levels in PBMCs were detected via quantitative real-time reverse transcription (qRT) PCR with SYBR Green (SYBR Premix Ex Taq II, Takara Bio Inc., Japan). qRT-PCRs were conducted using a QuantStudio 12K Flex Real-Time PCR System (Applied Biosystems, Foster City, CA, USA), and the cycle conditions of reactions were 95°C for 1 min, followed by 42 cycles at 95°C for 10 sec, 60°C for 30 sec, and 72°C for 1 min. Relative transcription levels of METTL3, METTL14, and WTAP were calculated via comparisons with the housekeeping gene (internal control) β-actin in the same sample, and the 2^-△△Ct method was used to express levels (25 (link)).

Zhang T.P., Li R., Wang L.J., Huang Q, & Li H.M. (2022). Roles of the m6A methyltransferases METTL3, METTL14, and WTAP in pulmonary tuberculosis. Frontiers in Immunology, 13, 992628.

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Profiling RNA-binding Proteins in PBMCs

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The PBMCs were isolated from 5 ml peripheral blood and stored at −80°C until processed. Total RNA was extracted from PBMCs using TRIzol Reagent (Invitrogen, Carlsbad, CA, USA), and the RNA concentration was detected with NanoDrop 2000 spectrophotometer (Thermo Scientific, USA). Next, the total RNA was reversely transcribed into cDNA by the PrimeScriptTM RT Reagent Kit (Takara Bio Inc., Japan).
In this study, the YTHDF1, YTHDF2, YTHDF3, YTHDC1, and YTHDC2 expression levels in PBMC were measured by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) with SYBR Green (SYBR Premix Ex Taq II, Takara Bio Inc., Japan), and this experiment was carried out in duplicate by using QuantStudio 12K Flex Real-Time PCR system (Applied Biosystems, Foster City, CA, USA). Thermal cycling conditions were as follows: 95°C for 1 min, followed by 42 cycles at 95°C for 10 s, 60°C for 30 s, and 72°C for 1 min. The relative expression levels of these genes were calculated by using the 2^−ΔΔCt method normalized to an endogenous control, and the housekeeping gene β-actin was used as an internal control in the same sample.

Li H.M., Tang F., Wang L.J., Huang Q., Pan H.F, & Zhang T.P. (2022). Association of N6-methyladenosine readers' genes variation and expression level with pulmonary tuberculosis. Frontiers in Public Health, 10, 925303.

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Quantifying CDX2 and VIL1 Expression

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Total RNA was extracted using TRI Reagent (Ambion, USA) and converted to cDNA using a PrimeScript RT reagent kit (Takara, Japan). Human CDX2 (forward 5′-TTCACTACAGTCGCTACATCACC-3′; reverse 5′-TTGTTGATTTTCCTCTCCTTTGC-3′) and VIL1 (forward 5′-GGCAAGAGGAACGTGGTAGC-3′; reverse 5′-CGGTCCATTCCACTGGATGA-3′) were amplified with SYBR Green (SYBR Premix Ex Taq II, Takara, USA) in a fluorescence reader ABI Prism 7500. The following PCR parameters were used: 95 °C for 30 seconds, 40 cycles of 95 °C for 15 seconds, 55 °C for 30 seconds and finally an elongation step at 72 °C for 30 seconds. Each reaction was performed in triplicate and normalized to GAPDH. Relative expression of the target genes was determined using the 2^−ΔΔCt method^{23 (link)}. Thereafter, expression was expressed as fold difference relative to that of the untreated control cells. The results are expressed as mean ± SD of representative triplicates.

Li Q., Zhu Y., Liu J., Yu X., Chen M., Dong N., Gong Y, & Yuan Y. (2017). HpSlyD inducing CDX2 and VIL1 expression mediated through TCTP protein may contribute to intestinal metaplasia in the stomach. Scientific Reports, 7, 2278.

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