7900 real time pcr system

Manufactured by Takara Bio

Sourced in China, Japan

The 7900 Real-time PCR System is a high-performance instrument designed for quantitative real-time PCR analysis. It features precise temperature control, sensitive detection, and advanced data analysis capabilities to support a wide range of real-time PCR applications.

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Close spelling variants of this product

ABI 7900 HT Fast Real-Time PCR System ABI 7900 HT Real-Time PCR system 7900 HT real-time PCR system ABI prism 7900 Real-Time PCR System ABI 7900 Real-Time PCR System 7900 Prism Real-Time PCR system Real‐Time System

6 protocols using 7900 real time pcr system

Quantitative Analysis of SPRED1 Gene Expression

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RNA was extracted from mononuclear cells of bone marrow samples using the TRIzol reagent (TaKaRa, Japan) and reverse transcribed using a PrimeScript Reverse Transcription Reagent Kit with gDNA Eraser (Perfect Real Time, TaKaRa) according to the manufacturer's protocol. The integrity of synthesized cDNA was confirmed using β-actin as the endogenous control.
Quantitative reverse transcription polymerase chain reaction (PCR) was performed using 7900 real-time PCR system and SYBR Green (TaKaRa) as a double-stranded DNA-specific dye. Target genes were amplified with primers designed by Invitrogen (Shanghai, China). Specific primer sequences of SPRED1 were as follows: forward: 5'-GATGAGCGAGAGACGGAGAC-3' and reverse: 5'-GTCTCTGAGTCTCTCCACGGA-3'. The following protocol was used for real-time PCR: 1 cycle at 95°C for 30 s, followed by 40 cycles at 95°C for 5 s and 60°C for 34 s, and then 1 cycle at 95°C for 15 s, 60°C for 1 min, 95°C for 15 s, and 60°C for 15 s. A melting curve was generated for every PCR amplicon to check the specificity of the PCR reaction. The relative level of SPRED1 was analyzed using the ABI 7900 Sequence Detection System (Applied Biosystems, CA, USA) and calculated using the 2^-ΔΔCt method.

Sun J., Zhang J., Wang Y., Li Y, & Zhang R. (2019). A Pilot Study of Aberrant CpG Island Hypermethylation of SPRED1 in Acute Myeloloid Leukemia. International Journal of Medical Sciences, 16(2), 324-330.

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Quantitative PCR for RNA Analysis

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Total RNA was obtained from the tissue samples, and cells were harvested using TRIzol kits (Invitrogen Life Technologies). Quantitative PCR was performed using an Applied Biosystems 7900 Real-time PCR System (Shanghai, China) and a TaqMan Universal PCR Master Mix (Takara, Dalian, China), according to the manufacturer’s instructions.

LIU J., TONG S.J., WANG X, & QU L.X. (2014). Farnesoid X receptor inhibits LNcaP cell proliferation via the upregulation of PTEN. Experimental and Therapeutic Medicine, 8(4), 1209-1212.

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Quantification of Src-1 and Twist1 mRNA

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Total RNA of tissue samples was extracted according to the operation protocol of TRIzol reagents (Invitrogen, California, USA). The quality and quantity of extracted RNA was evaluated by Nanodrop 2000 spectrophotometer. Complementary DNA was generated using RevertAid First Strand cDNA Synthesis kit (Thermo, Massachusetts, USA) and quantified using a standard SYBR-Green PCR kit protocol (Takara, Japan) in ABI 7900 Real Time PCR system. All samples were done in triplicate and normalized to GAPDH. The relative expression levels were calculated by the equation 2^−ΔΔCT. The primers for qRT-PCR were: Src-1: Forward: 3’-TCA CTT CAG TCC GCC ACT-5’; Reverse: 3’-TCG CCT GTT CCT GGT TGT-5’; Twist1: Forward: 3’-ACC ATC CTC ACA CCT CTG-5’; Reverse: 3’-GAT TGG CAC GAC CTC TTG-5’. The log2-transformed mRNA expression fold change of Src-1 and Twist1 when comparing tumor versus normal tissues was calculated by equation: Log2 (T/N) = log2 (Src-1 or Twist1 mRNA expression in tumor/ Src-1 or Twist1 mRNA expression in paired normal tissue).

Zhou J., Zhang J., Xu M., Ke Z., Zhang W, & Mai J. (2019). High SRC-1 and Twist1 expression predicts poor prognosis and promotes migration and invasion by inducing epithelial-mesenchymal transition in human nasopharyngeal carcinoma. PLoS ONE, 14(4), e0215299.

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Quantification of RNA Species in HASMCs

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Total RNA from treated HASMCs was isolated using TRIzol reagent (Invitrogen). For the quantification of lncRNA and mRNA, the cDNA was obtained by using PrimeScript RT reagent kit (Takara, Dalian, China). The real‐time PCR was performed on the 7900 Real‐time PCR System using SYBR Premix Ex Taq (Takara). For the quantification of miR‐150‐5p, cDNA was obtained by using Taqman MicroRNA Reverse Transcription kit (Thermo Fisher Scientific, Waltham, USA) and real‐time PCR was performed on the 7900 Real‐time PCR System using Taqman microRNA assay kit (Thermo Fisher Scientific). The fold changes for lncRNA, mRNA and miRNA expression were determined using comparative Ct method, and GAPDH was selected as the internal control for lncRNA and mRNA and U6 was selected as the internal control for miRNA.

Cai T., Cui X., Zhang K., Zhang A., Liu B, & Mu J. (2019). LncRNA TNK2‐AS1 regulated ox‐LDL‐stimulated HASMC proliferation and migration via modulating VEGFA and FGF1 expression by sponging miR‐150‐5p. Journal of Cellular and Molecular Medicine, 23(11), 7289-7298.

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RNA Extraction and qRT-PCR Analysis

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RNA was extracted from the tissue and cells using TRIzol (Invitrogen). cDNA was synthesized with HiScript 1^st Strand cDNA Synthesis Kit (Vazyme Biotech, China). For the quantification of mRNA and lncRNA, the cDNA was obtained by using PrimeScript RT reagent kit (Takara, Dalian, China). The real‐time PCR was performed by 7900 Real‐time PCR System using SYBR Premix Ex Taq (Takara). The primers using for quantitative RT-PCR analysis were listed in Table S1.

Ren Z., Hu Y., Sun J., Kang Y., Li G, & Zhao H. (2022). N6-methyladenosine methyltransferase WTAP-stabilized FOXD2-AS1 promotes the osteosarcoma progression through m6A/FOXM1 axis. Bioengineered, 13(4), 7963-7973.

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RNA Extraction and RT-qPCR Analysis

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Firstly, the total RNA in cancer cells was extracted by Trizol reagent (Invitrogen). Then, reverse transcriptions were performed as previously described. In brief, the extracted RNA was utilized to synthesize cDNA with the instructions of PrimerScript RT Reagent Kit (TaKaRa, Kyoto, Japan). The realtime PCR was performed according to SYBR Green PCR Master Mix (Applied Biosystems, Foster City, CA) on the 7900 Real-time PCR System using SYBR Premix Ex Taq (Takara). RT-qPCR primers were purchased from RiboBio Biotechnology Co., Ltd (Guangzhou, China), and listed in Supplementary Table S1. GAPDH was used as endogenous controls.

Liu D., Li Z., Zhang K., Lu D., Zhou D, & Meng Y. (2023). N⁶-methyladenosine reader YTHDF3 contributes to the aerobic glycolysis of osteosarcoma through stabilizing PGK1 stability. Journal of cancer research and clinical oncology, 149(8).

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