Ultrasybr mix

Manufactured by CWBIO

Sourced in China

UltraSYBR mix is a ready-to-use solution for quantitative real-time PCR (qPCR) applications. It contains all the necessary components, including a DNA-binding dye, buffer, and polymerase, to perform qPCR reactions.

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Lab products found in correlation

Trizol reagent, by Thermo Fisher Scientific (2 mentions) Takara reverse transcriptase kit, by Takara Bio (1 mentions) Reverse transcription kit, by CWBIO (1 mentions) Steponeplus real time pcr system, by Thermo Fisher Scientific (1 mentions) Rt pcr kit, by CWBIO (1 mentions) Primescript rt kit, by Takara Bio (1 mentions) Trizol reagent, by Takara Bio (1 mentions) Mir x mirna first strand synthesis, by Takara Bio (1 mentions) Quantstudio 6 pcr system, by Thermo Fisher Scientific (1 mentions) Primer premier software v 5, by Premier Biosoft (1 mentions) Cdna synthesis kit, by Vazyme (1 mentions)

Spelling variants of this product

2× SYBR UltraSYBR Mix (2 citations)

7 protocols using ultrasybr mix

Transcriptional Response to SAHA and PTX

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Total RNA isolation and cDNA synthesis from OC3/P cells treated with SAHA or PTX alone or in combination were performed using Trizol and the Takara Reverse Transcriptase kit (Takara, Otsu, Japan), respectively, according to the manufacturer’s instructions. The cDNA encoding the indicated genes was amplified with the following specific primers:
bcl-2, 5’-ATGTGTGTGGAGAGCGTCAAC-3’ and 5’-AGAGACAGCCAGGAGAAATCAAAC-3’; bax, 5’-GACGGCAACTTCAACTGGG-3’ and 5-CCTGGATGAAACCCTGAAGC-3’c-my-c, 5’-CTGCGACGAGGAGGAGAA-3’ and 5’-CCGAAGGAGAAGGGTGT-3’; mdr1, 5’-ATAATGCGACAGGAGATAGGC-3’ and 5’-ATGTTGCCATTGACTGAAAGAA-3’, actin, 5-TGGCACCCAGCACAATGAA-3 and 5-CTAAGTCATAGTCCGCCTAGAAGCA-3. Real-time PCR was performed using the UltraSYBR Mix (CWBIO, Beijing, China) with the Mx3000p Real-Time PCR System (Stratagene, La Jolla, CA, USA) using the following thermal cycling conditions: 10 min at 95°C followed by 40 cycles of 10 s at 95°C, 30 s at 60°C, and 30 s at 72°C. Data were analyzed according to the 2 ^–ΔΔCt method.

Liu Z., Tong Y., Liu Y., Liu H., Li C., Zhao Y, & Zhang Y. (2014). Effects of suberoylanilide hydroxamic acid (SAHA) combined with paclitaxel (PTX) on paclitaxel-resistant ovarian cancer cells and insights into the underlying mechanisms. Cancer Cell International, 14, 139.

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LPS-induced Fibroblast Transcription Assay

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Fibroblasts were treated with LPS, LPS＋hucMSC-ex, LPS＋hucMSC-NC-ex, and LPS＋hucMSC-si-ex. After 24 h, the cells were collected and lysed with TRIzol reagent (Invitrogen) to extract total cellular RNA. Complementary DNA (cDNA) was obtained using a reverse transcription kit (CWBIO). RT-PCR was performed using UltraSYBR mix (CWBIO). β-actin was used as the endogenous reference gene for all RT-PCR experiments. PCR primers are listed in Table 1.

Guo Q., Zhao Y., Li J., Huang C., Wang H., Zhao X., Wang M, & Zhu W. (2021). Galectin-3 Derived from HucMSC Exosomes Promoted Myocardial Fibroblast-to-Myofibroblast Differentiation Associated with β-catenin Upregulation. International Journal of Stem Cells, 14(3), 320-330.

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Real-Time qPCR of Gene Expression

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Real‐time Real‐time quantitative PCR (qPCR) was performed using an Applied Biosystems 7500 PCR machine with Ultrasybr mix (CW Bio). The cycling parameters were as follows: 2 min at 50°C, and 40 cycles of 15 s at 95°C and 1 min at 60°C. The relative mRNA expression level of each target gene is shown following normalization to the ACTB gene. The primer sequences are shown in Table S2.

Wang F., Cheng H., Zhang Q, & Guo J. (2023). Genetic mutations in ribosomal biogenesis gene TCOF1 identified in human neural tube defects. Molecular Genetics & Genomic Medicine, 11(5), e2150.

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Quantification of mRNA Expression

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Total RNA was extracted with the TRIzol reagent (Invitrogen). The cDNA was obtained using RT-PCR kits (CWBIO, Cambridge, MA, USA). The PCR program was conducted with UltraSYBR mix (CWBIO) on a StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). The mRNA expression was normalized to β-actin. All the sequences of PCR primers are listed in Table 1.

Li J., Xu X., Fei S., Wang R., Wang H., Zhu W, & Zhao Y. (2022). Small Extracellular Vesicles Derived from Human Umbilical Cord Mesenchymal Stem Cells Enhanced Proangiogenic Potential of Cardiac Fibroblasts via Angiopoietin-Like 4. Stem Cells International, 2022, 3229289.

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Gene Expression Analysis via qRT-PCR

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Samples were dissolved in TRIzol reagent (Takara, Japan), total RNA was extracted and the concentration was quantified. A total of 500 ng of RNA was reverse transcribed to cDNA using a Prime Script™ RT kit (Takara, Japan). cDNA was reverse transcribed to cDNA using Ultra SYBR mix (CWBIO, Beijing, China) and specific primers and then analyzed using a Bio-Rad IQ5 real-time analysis system (Bio-Rad, Hercules, CA, USA) with specific primers. The reaction mixture was predenatured at 95°C for 10 min, denatured at 95°C for 15 s, annealed at 60°C for 1 min and entered a melting-curve phase with 40 cycles of amplification. Relative expression levels were calculated using the 2-ΔΔCT method. Each reaction was performed in triplicate to obtain normalized expression levels of the target gene for GAPDH. For miRNA, 600 ng of RNA was transcribed into cDNA using a reverse transcription kit (Mir-X™ miRNA First-Strand Synthesis) available from Clontech. Real time polymerase chain reaction (RT–PCR) was performed using the miScript SYBR Green PCR kit and miRNA-specific primers. U6 was used as an internal control.

Xu C., Zhang H., Yang C., Wang Y., Wang K., Wang R., Zhang W., Li C., Tian C., Han C., Li M., Liu X., Wang Y., Li Y., Zhang J., Li Y., Luo L., Shang Y., Zhang L., Chen Y., Shen K, & Hu D. (2024). miR-125b-5p delivered by adipose-derived stem cell exosomes alleviates hypertrophic scarring by suppressing Smad2. Burns & Trauma, 12, tkad064.

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Quantification of Anthocyanin Biosynthesis Genes

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Total RNA was extracted from leaves at different developmental stages, as described above, and cDNA was synthesized from 2 μg total RNA using a cDNA synthesis kit (Vazyme, Nanjing, China) in a total volume of 20 μL. A total of nine DEGs related to anthocyanin biosynthesis in S0–S2 were selected for qRT-PCR. Gene-specific primers were designed using Primer Premier Software v.5.0 (Premier Biosoft, Palo Alto, CA, USA) (Table S3). Ultra SYBR Mix (CWBIO, Beijing, China) and the QuantStudio 6 PCR system (Thermo Fisher Scientific, Waltham, MA, USA) was used for qRT-PCR. Furthermore, 50 μL reaction mixture contained 2 μL cDNA (1:50 dilution), 25 μL of 2× Ultra SYBR Mix (CWBIO, Beijing, China), and 1 µL of each primer (100 nM final concentration). The amplification conditions were as follows: 95 °C for 10 min, 40 cycles of 95 °C for 15 s, and 60 °C for 1 min. A melting curve analysis (55–95 °C) was performed at 95 °C for 15 s, 60 °C for 1 min, 95 °C for 15 s, and 60 °C for 15 s to confirm the specificity of the PCR amplification. Actin gene was used as an internal control. Relative expression levels were calculated using the 2^−ΔΔCt method [66 (link)]. All experiments were performed using three independent samples from three independent biological replicates.

Zou J., Gong Z., Liu Z., Ren J, & Feng H. (2023). Investigation of the Key Genes Associated with Anthocyanin Accumulation during Inner Leaf Reddening in Ornamental Kale (Brassica oleracea L. var. acephala). International Journal of Molecular Sciences, 24(3), 2837.

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RNA Expression Analysis of MMS and TMZ Treatment

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After cells were grown to the exponential phase, the culture medium was aspirated, and fresh media containing MMS or TMZ were added. The cells were then incubated in a 37 °C CO₂ incubator for 30 min. After samples were washed with PBS, fresh DMEM medium was added to all samples for further incubation. The total RNA of all samples was isolated by using the Trizol method. Before reverse transcription, DNase I (DNase I, Ambion) was used to digest genomic DNA in the total RNA samples. Then, purified total RNA samples were detected with a UV–vis spectrophotometer to calculate the concentration, and reverse transcription was carried out using a reverse transcription kit. Real-time fluorescence quantitative PCR (qPCR) was performed using UltraSYBR mix (Cwbio) to measure the relative RNA level of tested genes. GAPDH was used as the negative control, and TransScript II Green One-Step RT-qPCR SuperMix (TransGen) was used for RT-qPCR. The primers used for qPCR are listed in the Supplementary Table S1.

Zuo Y., He J., Zhou Z., Sun J., Ouyang C., Huang H., Wang Y., Liu H, & Reed S.H. (2024). Long non-coding RNA LIP interacts with PARP-1 influencing the efficiency of base excision repair. Non-coding RNA Research, 9(3), 649-658.

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