Sybr rt pcr kit

Manufactured by Takara Bio

Sourced in China, Japan, United States, Switzerland

The SYBR RT-PCR kit is a reagent designed for real-time reverse transcription PCR (RT-PCR) analysis. It contains the necessary components for efficient reverse transcription and subsequent real-time PCR amplification using SYBR Green detection chemistry.

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Lab products found in correlation

Trizol reagent, by Thermo Fisher Scientific (50 mentions) Lightcycler, by Roche (15 mentions) Primescript rt reagent kit, by Takara Bio (8 mentions) Trizol reagent, by Takara Bio (7 mentions) Trizol, by Thermo Fisher Scientific (6 mentions) Rt reagent kit, by Takara Bio (5 mentions) Reverse transcriptase, by Takara Bio (5 mentions) Chromo 4, by Takara Bio (4 mentions) Rnaiso plus reagent, by Takara Bio (4 mentions) Lightcycler 480, by Roche (4 mentions) Taqman microrna reverse transcription kit, by Thermo Fisher Scientific (4 mentions) Abi 7500 real time pcr system, by Thermo Fisher Scientific (4 mentions) Cfx96 qpcr system, by Bio-Rad (3 mentions) Dna pcr kit, by Takara Bio (3 mentions) Rnase r, by Illumina (3 mentions) Primescript rt reagent kit with gdna eraser, by Takara Bio (3 mentions) Cfx96 real time pcr detection system, by Bio-Rad (3 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions) Stepone real time pcr system, by Thermo Fisher Scientific (2 mentions) Superscript 3 reverse transcriptase kit, by Thermo Fisher Scientific (2 mentions)

91 protocols using sybr rt pcr kit

Investigating Leaf Development in Rosa Multiflora

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The roots, stems, leaves, buds, petals, sepals, and leaves of the biennial cuttings of Rosa multiflora ‘Libellula’ were harvested to detect leaf development-related gene expressions. Five key stages of leaf development of Rosa multiflora ‘Libellula’ (P1–P5) were gathered to detect leaf development-related gene expressions. The newly growing leaves of the control and VIGS rose plants (three to four weeks old) were harvested for silencing effect identification. Total RNA was extracted using a SteadyPure Plant RNA Extraction Kit (Code: AG21019; Accurate Biotechnology Co., Ltd., Hunan, China); cDNA synthesis was performed with the RT reagent kit (Takara) according to the manufacturer’s protocol.
Real-time PCRs were obtained on a Chromo 4™ continuous fluorescence detector with the SYBR RT-PCR Kit (Takara), 20 µL reaction volume: 10 µL of SYBR Green I PCR mix, 0.5 µM forward and reverse primers, 1 µL of cDNA template and supplemented with the appropriate amount of ddH2O to 20 µL. Amplification conditions were: 2 min at 95 °C; 40 cycles of 15 s at 95 °C, 30 s at 58 °C, and 30 s at 72 °C. Fold changes of RNA transcripts were calculated by the 2^−∆∆Ct method [49 (link)]. We used RcUBC (LOC112195044) in R.Chinensis plants as the internal reference. The entire experiments were repeated three times, and primers used in the qPCR experiments are listed in Supplementary Table S1 (Table S1).

Xia M., Xu Q., Liu Y, & Ming F. (2022). Mutagenic Effect of 60Co γ-Irradiation on Rosa multiflora ‘Libellula’ and the Mechanism Underlying the Associated Leaf Changes. Plants, 11(11), 1438.

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Quantitative Analysis of RNA Expression

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Total RNA was isolated from cells using RNAiso Plus (TaKaRa Code: D9108A; TaKaRa Biotechnology Dalian Co., Ltd.), and the purity of the extracted RNA was determined at optical densities between 260 nm and 280 nm. The primers were as follows: IL-1β (GEO ID: NM_000576.2) primers, forward, 5′-TGATGGCTTATTACAGTGGCAATG-3′, reverse, 5′-GTAGTGGTGGTGGGA GATTCG-3′; TGF-β1 (GEO ID: NM_000660.5) primers, forward, 5′-GGAAACCCAC AACGAAATCTATG-3′, reverse, 5′-CGCCAGGAATTGTTGCTGTA-3′; COL1α1 (GEO ID: NM_000088.3) primers, forward, 5′-CCTCAAGGGCTCCAACGAG-3′, reverse, 5′-TCAATCACTGTCTTGCCCCA-3′; GAPDH (GEO ID: NM_002046.5) primers, forward, 5′-TGGTATCGTGGAAGGACTCATGAC-3′, reverse, 5′-ATGCCAGTGAGCTTCCCGTTCAGC-3′.
Quantitative RT-PCR (QRT-PCR) was conducted using SYBR^® RT-PCR kit (TaKaRa Code: DRR096A; TaKaRa Biotechnology Dalian Co., Ltd.), according to the manufacturer’s instructions.
QRT-PCR was performed with an iCycler iQ™ Real-Time PCR Detection System (Bio-Rad Laboratories Inc., Hercules, CA, USA) and universal cycling conditions (2 minutes at 50°C, 10 minutes at 95°C, 40 cycles of 15 seconds at 95°C, and 1 minute at 60°C). Dissociation curves were recorded after each run.

Wang R., Chen J., Zhang Z, & Cen Y. (2015). Role of chymase in the local renin-angiotensin system in keloids: inhibition of chymase may be an effective therapeutic approach to treat keloids. Drug Design, Development and Therapy, 9, 4979-4988.

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Quantitative RT-PCR for Gene Expression

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Total RNA was prepared from cells using TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. Complementary DNA (cDNA) was synthesized from 0.5 μg total RNA by reverse transcriptase (Takara, Dalian, China). Quantitative real-time RT-PCR (qRT-PCR) analysis was performed with the SYBR RT-PCR Kit (Takara, Dalian, China) and LightCycler (Roche Diagnostics, Indianapolis, IN) as described previously23 (link). Primers used for quantitative-PCR (qRT-PCR) amplification of β-Actin, and IL-1β were described previously23 (link). Data were normalized by the level of β-Actin expression in each sample.

Qin X., Jiang X., Jiang X., Wang Y., Miao Z., He W., Yang G., Lv Z., Yu Y, & Zheng Y. (2016). Micheliolide inhibits LPS-induced inflammatory response and protects mice from LPS challenge. Scientific Reports, 6, 23240.

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Quantitative Real-Time PCR Analysis of Gene Expression

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Total RNA was extracted from allografts using TRIzol reagent (Invitrogen, Carlsbad, CA, USA), according to the manufacturer’s instructions; cDNAs were synthesized using an oligo d(T) primer (Applied Biosystems, Foster City, California, U.S.) and a Superscript III Reverse Transcriptase Kit (Invitrogen). A StepOne™ Real-Time PCR System (Applied Biosystems) and a SYBR RT-PCR kit (Takara, Japan) were used for the quantitative real-time PCR analysis. All reactions were conducted in a 20 μl reaction volume in triplicate. The relative expression levels of a target gene were normalized to GAPDH expression. The specificity of qPCR was verified with a melting curve analysis and agarose gel electrophoresis. Primer sequences used in the RT-PCR analysis are shown in Table 1 [23 (link)].Table 1

RT-PCR primer sequence

Primer	Sequence
RORγt	Forward 5′-TCA CCT GAC CTA CCC GAG G-3′
RORγt	Reverse 5′-TCC AAG AGT AAG TTG GCC GTC-3′
GAPDH	Forward 5′-AGG TCG GTG TGA ACG GAT TTG-3′
GAPDH	Reverse 5′-TGT AGA CCA TGT AGT TGA GGT CA-3′
IL-17A	Forward 5′-TCG CCA TTC AGC AAG AAA TCC-3′
IL-17A	Reverse 5′-CAC AGG TGC AGC CAA CTT TTA-3′
IFN-γ	Forward 5′-GAA CTG GCA AAA GGA TGG TGA-3′
IFN-γ	Reverse 5′-TGT GGG TTG TTG ACC TCA AAC-3′
IL-4	Forward 5′-GGT CTC AAC CCC CAG CTA GT-3′
IL-4	Reverse 5′-GCC GAT GAT CTC TCT CAA GTG AT-3′
FOXP3	Forward 5′-TCA AGT ACC ACA ATA TGC GAC C-3′
FOXP3	Reverse 5′-CCA TCG GAT AAG GGT GGC A-3′

Ye Q., Zhang M., Wang Y., Fu S., Han S., Wang L, & Wang Q. (2017). Sirtinol regulates the balance of Th17/Treg to prevent allograft rejection. Cell & Bioscience, 7, 55.

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Quantitative PCR Analysis of Lung Tissue

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A total of 100 mg irradiated lung tissue was freshly isolated from each sample. Total RNA was isolated with TRIzol reagent (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA), and 1 mg total RNA from each sample was used for first-strand complementary DNA synthesis (37°C for 15 min; 85°C for 5 sec) with a RT-PCR kit (Takara Bio, Inc., Otsu, Japan). RT-qPCR (95°C for 1 min; 95°C for 10 sec; 58°C for 10 sec; 72°C for 10 sec; all for 40 cycles) was performed with the SYBR RT-PCR kit (Takara Bio, Inc.) on the Chromo4 Real-time PCR system (Bio-Rad Laboratories, Inc., Hercules, CA, USA). The level of GAPDH mRNA in each sample was used as an internal control. All reactions were performed in duplicate, and the results were analyzed by the 2^−ΔΔCq method (21 (link)). The primer sequences are stated in Table I.

Tang F., Li R., Xue J., Lan J., Xu H., Liu Y., Zhou L, & Lu Y. (2017). Azithromycin attenuates acute radiation-induced lung injury in mice. Oncology Letters, 14(5), 5211-5220.

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Quantitative Real-Time RT-PCR Analysis

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Total liver RNA was extracted using TRIzol (Takara, Tokyo, Japan) reagent according to the manufacturer’s instructions. The cDNA was synthesized using 1000 ng of total RNA in the first-strand cDNA synthesis reaction with PrimeScript RT reagent Kit (Takara). RT-PCR was performed using the CFX 96 q-PCR system (BIO-RAD, Hercules, CA, USA). A SYBR RT-PCR kit (Takara) were used for quantitative real-time RT-PCR analysis. All reactions were conducted in a 20 μl reaction volume in triplicate. The relative expression levels for a target gene were normalized by β-actin. Primers used for RT-PCR analysis are:
Gclc forward: 5′- ATGTGGACACCCGATGCAGTATT
Gclc reverse: 5′-TGTCTTGCTTGTAGTCAGGATGGTTT
Gclm forward: 5′- TGGAGCAGCTGTATCAGTGG
Gclm reverse: 5′- AGAGCAGTTCTTTCGGGTCA
Gstm1 forward: 5′- CTACCTTGCCCGAAAGCAC
Gstm1 reverse: 5′- ATGTCTGCACGGATCCTCT
Nqo1 forward: 5′- AGCGTTCGGTATTACGATCC
Nqo1 reverse: 5′- AGTACAATCAGGGCTCTTCTCG
Il1b forward: 5′- TGTAATGAAAGACGGCACACC
Il1b reverse: 5′- TCTTCTTTGGGTATTGCTTGG
Tnf forward: 5′- TTCTATGGCCCAGACCCTCA
Tnf reverse: 5′- TTTGCTACGACGTGGGCTAC
Il6 forward: 5′- GCTACCAAACTGGATATAATCAGGA
Il6 reverse: 5′- CCAGGTAGCTATGGTACTCCAGAA
Cxcl-10 forward: 5′- GCTGCCGTCATTTTCTGC
Cxcl-10 reverse: 5′- TCTCACTGGCCCGTCATC
Actb forward: 5′- TGACAGGATGCAGAAGGAGA
Actb reverse: 5′- ACCGATCCACACAGAGTACT.

Xu D., Chen L., Chen X., Wen Y., Yu C., Yao J., Wu H., Wang X., Xia Q, & Kong X. (2017). The triterpenoid CDDO-imidazolide ameliorates mouse liver ischemia-reperfusion injury through activating the Nrf2/HO-1 pathway enhanced autophagy. Cell Death & Disease, 8(8), e2983-.

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Isolation and Quantification of Lung Fibroblast RNA

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After different treatment, total RNA was isolated from the primary lung fibroblast using Trizol Reagent (Sigma, St. Louis, MO, USA) according to the manufacturer’s instructions. Then, RNA was reverse transcribed into cDNA using a reverse transcription reaction system (Takara Bio Inc., Dalian, China). RT-qPCR was performed with the SYBR RT-PCR kit (Takara Bio Inc.). The primer sequences are shown in Table 1.

Zhang Y., Du H., Yu X, & Zhu J. (2020). Fucoidan attenuates hyperoxia-induced lung injury in newborn rats by mediating lung fibroblasts differentiate into myofibroblasts. Annals of Translational Medicine, 8(22), 1501.

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Molecular Mechanisms of Cell Apoptosis

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Anti-Parp-1 (ab74290), anti-cleaved-Parp-1 (ab32064), anti-Caspase 3 (ab13847), anti-cleaved-Caspase 3 (ab2302), anti-β-actin, anti-APAF1 (ab2001), anti-pan-AKT (ab8805), anti-HSP90-beta (ab203085), anti-Ki67 (ab16667), anti-Caspase 9 (ab184786), anti-FKHR (ab179540), anti-IKK-beta (ab124957), anti-p70S6K (ab32529), anti-P65 (ab16502), anti-HSP70 (ab5439), anti-HSC70 (ab51052), anti-IgG (ab133470), anti-HA (ab236632), anti-His (ab18184), Dactinomycin and ADP/ATP Ratio Assay Kits were purchased from Abcam (Britain). TRIzol reagent was acquired from Invitrogen (USA); the SYBR RT‒PCR Kit and DNA PCR kit were from Takara Bio Inc. (Japan); RNase R was from Epicenter (USA).
Primers and siRNAs were designed and synthesized by Sangon Biotech (China). The ATP Assay Kit was purchased from Beyotime (China). The dual-luciferase reporter assay system was obtained from Promega (USA).

Deng Y., Zeng X., Lv Y., Qian Z., Guo P., Liu Y, & Chen S. (2023). Cdyl2-60aa encoded by CircCDYL2 accelerates cardiomyocyte death by blocking APAF1 ubiquitination in rats. Experimental & Molecular Medicine, 55(4), 860-869.

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Quantitative Real-Time PCR for Gene Expression

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RNA was isolated from breast cancer tissue and cell lines by using TRIzol Reagent (Invitrogen) according to the manufacturer’s protocol, as previously described.⁴¹ (link) RNA samples were reverse transcribed to cDNA, and the relative transcript abundances were then estimated by quantitative real-time PCR using a SYBR RT-PCR kit (Takara, Shiga, Japan) according to the manufacturer’s instructions. The 2^-ΔΔCt method was used to calculate the relative gene expression levels, which were normalized to those of beta-actin. The primer sequences are listed in Table 1.Table 1

Sequences of Primers Used in qPCR Experiments

Gene	Primers sequences
Human β-actin	Forward: 5′-AGCGAGCATCCCCCAAAGTT-3′
Human β-actin	Reverse: 5′-GGGCACGAAGGCTCATCATT-3′
Human GPR1	Forward: 5′-GGAGCTCAGCATTCATCACA-3′
Human GPR1	Reverse: 5′-GACAGGCTCTTGGTTT CAGC-3′

Huang C., Dai X.Y., Cai J.X., Chen J., Wang B.B., Zhu W., Wang E., Wei W, & Zhang J.V. (2020). A Screened GPR1 Peptide Exerts Antitumor Effects on Triple-Negative Breast Cancer. Molecular Therapy Oncolytics, 18, 602-612.

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RNA Extraction and Expression Analysis

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The TRIzol reagent (Invitrogen, Carlsbad, CA, USA) was used to isolate total RNA, including microRNA, according to the manufacturer’s instructions. For mRNA expression, cDNA synthesis was performed with Oligo (dT), and a SYBR RT-PCR kit (Takara, Otsu, Japan) was used for mRNA quantification with specific primers. For the detection of Cdr1as expression, specific primers were used (Forward: 5’-GTGTCTCCAGTGTATCGGCG-3’; Reverse: 5’-TACTGGCACCACTGGAAACC-3’) as described before [26 (link)]. For miR-7a expression, microRNA was analyzed by using the TaqMan MicroRNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA, USA) with provided RT-U6 and microRNA-specific stem-loop primers, and the expression levels were determined through TaqMan MicroRNA assays with the TaqMan Universal PCR Master Mix (Applied Biosystems). All reactions were preceded on the ABI 7500 real-time PCR system (Applied Biosystems) by standard protocols. The comparative threshold cycle (Ct) value was calculated and analyzed by using the 2^-ΔΔCT method, with U6 and β-actin as an internal control.

Geng H.H., Li R., Su Y.M., Xiao J., Pan M., Cai X.X, & Ji X.P. (2016). The Circular RNA Cdr1as Promotes Myocardial Infarction by Mediating the Regulation of miR-7a on Its Target Genes Expression. PLoS ONE, 11(3), e0151753.

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