Dynamo cdna synthesis kit

Manufactured by New England Biolabs

Sourced in United States

The DyNAmo cDNA Synthesis Kit is a product offered by New England Biolabs. It is designed for the reverse transcription of RNA to complementary DNA (cDNA).

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Lab products found in correlation

Trizol reagent, by Thermo Fisher Scientific (4 mentions) Sybr green, by Thermo Fisher Scientific (2 mentions) Superreal premix, by Tiangen Biotech (1 mentions) Maxima first strand cdna synthesis kit, by Thermo Fisher Scientific (1 mentions) C1000 thermal cycler, by Bio-Rad (1 mentions) Sybr green, by Quanta Biosciences (1 mentions) 35s methionine, by GE Healthcare (1 mentions) Sybr green mix, by Bio-Rad (1 mentions) γ 32p atp, by GE Healthcare (1 mentions) Protein a magnetic beads, by New England Biolabs (1 mentions) Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions) Dnase, by Promega (1 mentions)

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CDNA synthesis kit (5 citations)

6 protocols using dynamo cdna synthesis kit

RNA Extraction and qRT-PCR Quantification

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Total RNA was extracted by the TRIzol reagent (Life Technologies) according to the manufacturer's instructions. Total RNA concentration was estimated by absorbance at 260 nm using an ultraviolet spectrophotometer, and its integrity was confirmed by denaturing agarose gel electrophoresis. The first-strand cDNA generation was performed with a DyNAmo cDNA Synthesis Kit (New England Biolabs). Quantitative real-time PCR (qRT-PCR) was carried out with SuperReal PreMix (Tiangen Biotech) in triplicate times. The relative expression level was normalized by internal reference β-actin through the 2-^∆∆Ct method.
qRT-PCR primers for PTK7 are sense, 5′- catggtaccgttgtatgagca-3′, and antisense, 5′- ttgtctgtcacccactctgg-3′.
qRT-PCR primers for β-actin are sense, 5′ -tcccggcatgtgcaaggcc-3′, and antisense, 5′-catctcttgctcgaagtcca-3′.

Sun J.J., Li H.L., Guo S.J., Ma H., Liu S.J., Liu D, & Xue F.X. (2019). The Increased PTK7 Expression Is a Malignant Factor in Cervical Cancer. Disease Markers, 2019, 5380197.

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RNA Isolation and qPCR Analysis

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RNA was isolated using TRIzol reagent (Invitrogen) following the manufacturer's specifications. cDNA was synthesized using either the DyNAmo cDNA synthesis kit (New England Biolabs, F470) or the Maxima First Strand cDNA synthesis kit (Thermo, K1642). qPCR was performed using SYBR Green (Applied Biosystems).

Minas T.Z., Surdez D., Javaheri T., Tanaka M., Howarth M., Kang H.J., Han J., Han Z.Y., Sax B., Kream B.E., Hong S.H., Çelik H., Tirode F., Tuckermann J., Toretsky J.A., Kenner L., Kovar H., Lee S., Sweet-Cordero E.A., Nakamura T., Moriggl R., Delattre O, & Üren A. (2016). Combined experience of six independent laboratories attempting to create an Ewing sarcoma mouse model. Oncotarget, 8(21), 34141-34163.

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RNA Isolation and qRT-PCR Analysis

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RNA was isolated 5 days after lentiviral infection and puromycin selection with Trizol reagent (Invitrogen) following the manufacturer's protocol. cDNA was synthesized with DyNAmo cDNA synthesis kit (F470; New England Biolabs), and qRT-PCR was carried out in triplicate by SYBR Green (Quanta Biosciences) using a C1000 Thermal Cycler (BioRad). See Supplementary Methods for primer sequences.

Chen R., Khatri P., Mazur P.K., Polin M., Zheng Y., Vaka D., Hoang C.D., Shrager J., Xu Y., Vicent S., Butte A, & Sweet-Cordero E.A. (2014). A meta-analysis of lung cancer gene expression identifies PTK7 as a survival gene in lung adenocarcinoma. Cancer research, 74(10), 2892-2902.

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Quantitative Analysis of Gene and miRNA Expression in MEF and KLA Cells

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RNA was analyzed as previously described (82 (link)). MEFs were treated for 72 hours with adenoviruses and grown in fresh medium for an additional 72 hours. Then, MEFs were plated and RNA harvested after 24 hours with TRIzol reagent (Invitrogen). KLA cells were treated for 48 hours with adenoviruses and then fresh medium was added for 24 hours before RNA isolation. cDNA was synthesized with a DyNAmo cDNA synthesis kit (F470, New England Biolabs) and qRT-PCR was performed using SYBR Green (Applied Biosystems). GAPDH and HPRT were used as housekeeping genes. For miRNA analyses, TaqMan assays were used (miR181a, ID 000480; miR181b, ID 001098). RNU6 and Sno135 were reference genes in mouse cell lines. RNU6 and RNU48 were used as housekeeping genes in human cell lines.

Valencia K., Erice O., Kostyrko K., Hausmann S., Guruceaga E., Tathireddy A., Flores N.M., Sayles L.C., Lee A.G., Fragoso R., Sun T.Q., Vallejo A., Roman M., Entrialgo-Cadierno R., Migueliz I., Razquin N., Fortes P., Lecanda F., Lu J., Ponz-Sarvise M., Chen C.Z., Mazur P.K., Sweet-Cordero E.A, & Vicent S. (2020). The Mir181ab1 cluster promotes KRAS-driven oncogenesis and progression in lung and pancreas. The Journal of Clinical Investigation, 130(4), 1879-1895.

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Radioactive Labeling Reagents Protocol

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[γ-³²P]ATP and [³⁵S]methionine were purchased from GE Healthcare (Munich), and ³H₂O and [¹⁴C]sucrose were obtained from Biotrend (Cologne). SYBR Green Mix was from BioRad (Munich), the DyNAmo™ cDNA Synthesis Kit and Protein A magnetic beads were from New England Biolabs (Frankfurt am Main). Goat anti-(rabbit IgG)-alkaline phosphatase was obtained from Biomol (Hamburg). Purified KdpFABC protein was supplied by Marc Bramkamp (Osnabrück University). RNase-free deoxyribonuclease I was from Fermentas (St. Leon-Rot), and silicone oil (DC550) was from Serva (Heidelberg). All other reagents were reagent grade and obtained from commercial sources.

Heermann R., Zigann K., Gayer S., Rodriguez-Fernandez M., Banga J.R., Kremling A, & Jung K. (2014). Dynamics of an Interactive Network Composed of a Bacterial Two-Component System, a Transporter and K+ as Mediator. PLoS ONE, 9(2), e89671.

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Leaf RNA Extraction and cDNA Synthesis

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Total RNA was extracted from 150 mg of leaf tissue using RNeasy plant mini RNA kit (50) (Qiagen, Valencia, CA), and RNA samples were further treated with DNase (Promega, Madison, WI, USA) to eliminate DNA contamination. RNA Integrity was confirmed by agarose gel electrophoresis using the Agilent 2100 Bioanalyzer (Agilent Technologies, Inc., Santa Clara, CA). cDNA was synthesized using the DyNAmo cDNA Synthesis Kit (New England Biolabs, Ipswich,MA, USA). The qRT-PCR analysis primer pairs of the corresponding genes were designed according to sequences obtained from the Phytozyme website (Table 2). 20 μl reaction including 15 ng of random hexamers, 10 units of Moloney murine leukemia virus (M-MuLV) RNase H + reverse transcriptase (RT) solution, and appropriate buffer containing dNTPs and MgCl 2 in a final concentration of 5 mM (1×), The synthesis reaction lasted 10 min at 25°C, followed by 30 min at 37°C, 5 min at 85°C and 5 min at 4°C. Each RNA sample had two replicate RT reactions.

Liu Y.M., Hu G., WU G., Liu Y., Zhao B., & Zhang X.Z. (2020). Differential responses of antioxidants and dehydrin in two Switchgrass (Panicum virgatum L.) cultivars contrasting in drought tolerance. Tropical Plant Research, 7(1), 255-267.

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