Revertra ace qpcr rt kit

Manufactured by Qiagen

Sourced in Germany

The ReverTra Ace qPCR RT Kit is a reagent kit designed for reverse transcription and real-time quantitative PCR (RT-qPCR) analysis. The kit includes components necessary for cDNA synthesis and subsequent qPCR amplification.

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Lab products found in correlation

Trizol, by Thermo Fisher Scientific (4 mentions) Rneasy mini kit, by Qiagen (3 mentions) Propidium iodide, by Beyotime (1 mentions) Bcl 2, by Abcam (1 mentions) Gapdh, by Abcam (1 mentions) Bcl2 associated x (bax), by Abcam (1 mentions) Cyclin b1, by Cell Signaling Technology (1 mentions) Fitc annexin 5 apoptosis detection kit, by BD (1 mentions) P21 waf1 cip1, by Cell Signaling Technology (1 mentions) Cdc25a, by Cell Signaling Technology (1 mentions) Phosphor cdc2, by Cell Signaling Technology (1 mentions) Pifithrin α pft α, by Beyotime (1 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Dulbecco s modified eagle medium dmem, by Thermo Fisher Scientific (1 mentions) Geneticin g 418, by Solarbio (1 mentions) Kod sybr qpcr mix, by Toyobo (1 mentions)

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RT-qPCR kit (5 citations) QPCR kits (2 citations)

8 protocols using revertra ace qpcr rt kit

Actinomycin-Induced Lung Cancer Apoptosis

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Actinomycin V (>98%) and actinomycin D (>98%) were dissolved and their concentration adjusted by dimethyl sulfoxide (DMSO) when required while the control group was treated with DMSO only. MTT and DAPI were acquired from Sigma-Aldrich Crop. (St. Louis, MO, USA). Annexin V-FITC Apoptosis Detection Kit was obtained from BD Biosciences (San Jose, CA, USA). Phosphor-Cdc2, Cdc2, Cdc25A, Cyclin B1, p53 and p21^Waf1/Cip1 were supplied by Cell Signaling Technology (CST, Inc, Beverly, MA, USA). Bcl-2, Bax and GAPDH antibodies were supplied by Abcam, Inc. (Cambridge, MA, USA). Propidium iodide and Pifithrin-α (PFT-α) were purchased from Beyotime Institute of Biotechnology (Shanghai, China). RNeasy mini kit and Rever Tra Ace qPCR RT kit were purchased from QIAGEN (Hilden, Germany) and Toyobo (Osaka Prefecture, Japan), respectively.
Human non-small-cell lung carcinoma cell lines A549, NCI-H1299 (p53-deficient) and normal human bronchial epithelial cells BEAS-2B were obtained from the Shanghai Institute for Biological Sciences (SIBS), Chinese Academy of Sciences (Shanghai, China) and we cultured the cells according to the supplier’s instructions.

Lin S.Q., Jia F.J., Zhang C.Y., Liu F.Y., Ma J.H., Han Z., Xie W.D, & Li X. (2019). Actinomycin V Suppresses Human Non-Small-Cell Lung Carcinoma A549 Cells by Inducing G2/M Phase Arrest and Apoptosis via the p53-Dependent Pathway. Marine Drugs, 17(10), 572.

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Cytotoxicity Assay of Anticancer Drugs

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Roswell Park Memorial Institute (RPMI)-1640 medium, Dulbecco’s Modified Eagle Medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco (Gaithersburg, MD, USA). MTT, DOX, MITX, VCR, FTC, VRP, cisplatin, and the PREDEASY™ ATPase Kit were purchased from Sigma Corp. (St. Louis, MO, USA). Geneticin (G418) was purchased from Solarbio Science and Technology Co., Beijing, China. Monoclonal antibodies against glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ABCB1, and ABCG2 were obtained from Santa Crus Biotechnology (Dallas, TX, USA). RNeasy mini kit and Rever Tra Ace qPCR RT kit were purchased from QIAGEN (Hilden, Germany) and Toyobo (Osaka Prefecture, Japan), respectively. LS-2-3j, MTT, DOX, MITX, VCR, FTC, VRP, and cisplatin were dissolved in dimethylsulfoxide (DMSO) as a 10 mmol/L solution, stored at −20 °C.

Guo C., Liu F., Qi J., Ma J., Lin S., Zhang C., Zhang Q., Zhang H., Lu R, & Li X. (2018). A Novel Synthetic Dihydroindeno[1,2-b] Indole Derivative (LS-2-3j) Reverses ABCB1- and ABCG2-Mediated Multidrug Resistance in Cancer Cells. Molecules, 23(12), 3264.

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Quantitative Analysis of Plasma Cell Markers

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BM samples were respectively acquired from MM patients at diagnosis and from health donors at donation, which were then submitted to isolate plasma cells using CD138-immunomagnetic beads (cat. no. 130–051-301, Miltenyi Biotec, Paris, France) in accordance with product manual. Subsequently, quantitative analysis of EGFL7, ITGB3, and KLF2 expression was carried out by reverse transcription quantitative polymerase chain reaction (RT-qPCR). The detailed processes of RT-qPCR were as follows: (1) total RNA was extracted using RNeasy Protect Mini Kit (cat. no. 74124, Qiagen, Duesseldorf, Nordrhein-Westfalen, Germany). (2) The RNA was conversed to cDNA using ReverTra Ace® qPCR RT Kit (cat. no. FSQ-201, Toyobo, Osaka, Kansai, Japan). (3) The quantitative qPCR was carried out using KOD SYBR® qPCR Mix (cat. no. 7772383, Toyobo, Osaka, Kansai, Japan). (4) The PCR amplification was conducted as follows: 95 °C for 3 min, 40 cycles of 95 °C for 5 s, 61 °C for 10 s, 72 °C for 30 s. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was served as the internal reference. The calculation of mRNA expressions was according to 2^−ΔΔCt formula [15 (link)]. Primer design was conducted with a previous study used as reference [9 (link)].

Li Y., Zhang L, & Gong J. (2021). Relation among EGFL7, ITGB3, and KLF2 and their clinical implication in multiple myeloma patients: a prospective study. Irish Journal of Medical Science, 191(5), 1995-2001.

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Quantifying ABC Transporter Expression

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Real-time PCR was carried out to analyze the mRNA levels of ABCB1 and ABCG2. Cells were incubated in 6-well plates and allowed to become adherent overnight. Then, the cells were exposed to the test compound LS-2-3j (0, 0.25, 0.5 and 1 μmol/L) for 24 or 48 h. After that, RNA was extracted by the RNeasy Mini Kit (QIAGEN) according to the manufacturer’s instructions. Then, cDNA was synthesized through reverse transcription using the Rever Tra Ace qPCR RT kit (QIAGEN) and according to the related instructions. PCR amplification was performed in an 8-tube strip format (Axygen, Union City, CA, USA) in triplicate. For each reaction, 1 μL SYBR Green PCR Master Mix (1×), 1 μL forward and reverse primer (10 μmol/L), and 1 μL template cDNA were added to a final volume of 20 μL with diethyl pyrocarbonate (DEPC) water. Primers were based on the ABCB1 gene (forward: 5′-CCATGGAGAAGGCTGGGG-3′, reverse: 5′-CCAAGTTGTCATGGATGACC-3′), the ABCG2 gene (forward: 5′-GCCATAGCAGCAGGTCAG-3′, reverse: 5′-AGCCGTAAATCCATATCGTG-3′), and the GAPDH gene (forward: 5′-GACCTGACCTGCCGTCTA-3′, reverse: 5′-AGGAGTGGGTGTCGCTGT-3′). Reactions were performed for 45 cycles of sequential denaturation at (95 °C, 2 min), annealing (58 °C, 15 s) and extension (72 °C, 20 s). All primers in the above experiments were synthesized by Rui Bochenko Biological Technology Co., Ltd. (Qingdao, China).

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Prefrontal Cortex Dissection and RNA Extraction

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Twenty-four hours after the behavioral experiment, rats were anesthetized by pentobarbital (70 mg/kg, i.p.) and then killed by decapitation. PFCs were dissected (+5.16 mm to 3.00 mm from Bregma) according to the stereotactic atlas of Paxinos and Watson. Frozen PFCs were thawed in TRIzol (Invitrogen, Cat. No. 15596018) and then processed according to the manufacturer's protocol. Total RNA was reverse-transcribed using the ReverTra Ace qPCR RT Kit (Qiagen, Cat. No. 205313).

Li H., Chen J., Ding Q., Lu G., Wu N., Su R., Li F., & Li J. (2020). Behavioral Sensitization Induced by Methamphetamine Causes Differential Alterations in Gene Expression and Histone Acetylation of the Prefrontal Cortex in Rats.

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Postmortem Prefrontal Cortex Analysis

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Twenty-four hours after the behavioral experiment, rats were anesthetized with pentobarbital (70 mg/kg, i.p.) and then euthanized by decapitation. PFC tissues were dissected (+5.16 mm to 3.00 mm from the bregma) according to the stereotactic atlas published by Paxinos and Watson. Frozen PFC tissues were thawed in TRIzol (Invitrogen, Cat. No. 15596018) and then processed according to the manufacturer's protocol. Total RNA was reverse-transcribed using the ReverTra Ace qPCR RT Kit (Qiagen, Cat. No. 205313).

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Rat PFC RNA Extraction Protocol

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Twenty-four hours after behavioral experiment, rats were anesthetized by pentobarbital (70 mg/kg, i.p.) and then killed by decapitation. PFCs were dissected according to the stereotactic atlas of Paxinos and Watson. Frozen PFCs were thawed in TriZol (Invitrogen, Cat.15596018) and processed according to the manufacturer's protocol. Total RNA was reverse-transcribed using ReverTra Ace qPCR RT Kit (Qiagen, Cat.205313).

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Postmortem Prefrontal Cortex Analysis

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Li H., Chen J., Ding Q., Lu G., Wu N., Su R., Li F., & Li J. (2021). Behavioral Sensitization Induced by Methamphetamine Causes Differential Alterations in Gene Expression and Histone Acetylation of the Prefrontal Cortex in Rats.

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