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High capacity dna reverse transcription kit

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The High capacity DNA reverse transcription kit is a laboratory tool designed for the reverse transcription of RNA to complementary DNA (cDNA). It enables the conversion of RNA samples into cDNA for further analysis or applications.

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Lab products found in correlation

Power sybr green pcr master mix, by Thermo Fisher Scientific (8 mentions) Trizol reagent, by Thermo Fisher Scientific (7 mentions) Taqman master mix, by Thermo Fisher Scientific (4 mentions) Rneasy mini kit, by Qiagen (3 mentions) Stepone real time pcr system, by Thermo Fisher Scientific (2 mentions) Steponeplus real time pcr system, by Thermo Fisher Scientific (2 mentions) 7500 fast real time pcr system, by Thermo Fisher Scientific (1 mentions) Isolate rna mini kit, by Meridian Bioscience (1 mentions) Genelute mammalian total rna miniprep kit, by Merck Group (1 mentions) 7300 real time pcr system, by Thermo Fisher Scientific (1 mentions) Ab7500, by Thermo Fisher Scientific (1 mentions) Ambion rnaqueous micro kit, by Thermo Fisher Scientific (1 mentions) Rneasy plant mini kit, by Qiagen (1 mentions) Taqman gene expression assay, by Thermo Fisher Scientific (1 mentions) Lightcycler 96 system, by Roche (1 mentions) Lightcycler 96, by Roche (1 mentions) Biodrop duo, by Harvard Bioscience (1 mentions) Qubit fluorometer, by Thermo Fisher Scientific (1 mentions) Rneasy micro isolation kit, by Qiagen (1 mentions) Stepone plus equipment, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Reverse transcription kit (1270 citations) High Capacity Kit (98 citations) High-capacity complementary DNA reverse transcription kit (89 citations) High-capacity complementary DNA (cDNA) reverse transcription kit (9 citations) Applied Biosystems high-capacity complementary DNA reverse transcription kit (4 citations) Transcription Kits (2 citations) High-capacity copy DNA Reverse Transcription Kit (2 citations)

17 protocols using high capacity dna reverse transcription kit

1

Arabidopsis RNA Extraction and RT-qPCR Analysis

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Total Arabidopsis RNA was isolated according to the manufacturer's protocol using an Isolate RNA Minikit (Bioline, NJ). cDNA was synthesized using a high‐capacity DNA reverse transcription kit (Applied Biosystems, CA). RT‐qPCR reactions were done using a 7500 Fast Real‐Time PCR system (Applied Biosystems, CA) as described earlier (Mooney et al., 2019 (link)). Arabidopsis Actin2 (At3g18780) was used as the internal control gene, and all experiments were repeated at least three times as biological replicates, if not otherwise stated. For in planta detection of U‐PEST and U‐SBC expression specific primer pairs were designed that covered eK and PEST or SBC regions, respectively. All primers used are listed in Table S1. The 2(‐Delta Delta C(T)) method was used to calculate relative gene expression (Livak & Schmittgen, 2001 (link)).
Al‐Saharin R., Mooney S., Dissmeyer N, & Hellmann H. (2022). Using CRL3BPM E3 ligase substrate recognition sites as tools to impact plant development and stress tolerance in Arabidopsis thaliana. Plant Direct, 6(12), e474.
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2

Quantitative Analysis of mRNA Expression

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Total RNA was isolated from homogenized tissues (rat aorta, porcine and bovine coronary arteries) using the GenElute™Mammalian Total RNA Miniprep Kit (Sigma) including DNAse treatment of samples. cDNA was synthesized using the High Capacity DNA Reverse Transcription Kit (Applied Biosystems, Vienna, Austria). Primers for mRNA expression analysis were designed on the basis of published rat, porcine and bovine nucleic acid sequences of GenBank (NCBI) using the Primer-BLAST software (Table 1). Amplification efficiency of the primers was determined by qPCR analysis using serial dilutions of the cDNA template. Efficiency was calculated from the slope of the curve using the following equation: E = 10(−1/slope) (Table 1). Real-time PCR analysis was performed with ∼10 ng cDNA and primer concentrations of 100 nM using Power SYBR® Green PCR Master Mix (Applied Biosystems, Vienna, Austria). Reactions were carried out on a 7300 Real-Time PCR System (Applied Biosystems, Vienna, Austria). Cycling conditions were as follows: 2 min at 50 °C, 10 min at 95 °C, 50 cycles of 15 s at 95 °C, and for 1 min at 60 °C. Melting curve analysis consisted of the following steps: 15 s at 95 °C, 30 s at 60 °C, and 15 s at 95 °C. Data were analyzed and calculated according to Pfaffl [29] . Data were calculated relative to rat aortic ALDH2 mRNA expression after normalization to cyclophilin A.
Neubauer R., Wölkart G., Opelt M., Schwarzenegger C., Hofinger M., Neubauer A., Kollau A., Schmidt K., Schrammel A, & Mayer B. (2015). Aldehyde dehydrogenase-independent bioactivation of nitroglycerin in porcine and bovine blood vessels. Biochemical Pharmacology, 93(4), 440-448.
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3

Quantitative RT-PCR Gene Expression Analysis

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The total RNA was extracted using Trizol reagent according to manufacturer’s protocols (Invitrogen). 2 μg of RNA was used for reverse transcription using high capacity DNA reverse transcription kit (Applied Biosystems Cat No: 4368813) and qRT-PCR was performed using Taqman master mix or Power SYBR green PCR master mix (Applied biosystems cat No: 4367659) according to manufacturer’s protocols on Step One Real-Time PCR system (Applied Biosystems). The following cycling program was used for SYBR green assay: initial denaturation at 95°C for 10 min; 40 cycles of 95°C for 15 s and 60°C for 60 s; followed by a melt curve step. For Taqman assay: Initial denaturation 50°C for 2 min and 95°C for 10 min; 40 cycles of 95°C for 15 s and 60°C for 60 s. mRNA expression profiles were normalized to levels of housekeeping gene Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) or β-Actin in each sample and the fold-change in expression was calculated by the 2−ΔΔCt method.
Mehto S., Jena K.K., Nath P., Chauhan S., Kolapalli S.P., Das S.K., Sahoo P.K., Jain A., Taylor G.A, & Chauhan S. (2019). The Crohn’s Disease Risk Factor IRGM Limits NLRP3 Inflammasome Activation by Impeding Its Assembly and by Mediating Its Selective Autophagy. Molecular Cell, 73(3), 429-445.e7.
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4

Quantitative RT-PCR Analysis of Viral RNA

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RNA was extracted from cells using TRIZOL by following the manufacturer’s protocol. Viral RNA from the supernatant was extracted by an automated RNA isolation system. The cDNA was synthesized using the high capacity DNA reverse transcription kit (Applied Biosystems,#4368813), and qRT–PCR was performed using TaqMan master mix (Applied Biosystems, #4369016) or Power SYBR green PCR master mix (Applied Biosystems, #4367659) according to the manufacturer's protocol. For normalization of the assay, the housekeeping gene GAPDH or β‐Actin was used. The fold change in expression was calculated by the 2−ΔΔCt method. Excel or GraphPad is used for generating graphs and statistics.
Nath P., Chauhan N.R., Jena K.K., Datey A., Kumar N.D., Mehto S., De S., Nayak T.K., Priyadarsini S., Rout K., Bal R., Murmu K.C., Kalia M., Patnaik S., Prasad P., Reggiori F., Chattopadhyay S, & Chauhan S. (2021). Inhibition of IRGM establishes a robust antiviral immune state to restrict pathogenic viruses. EMBO Reports, 22(11), e52948.
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5

Quantitative RNA Expression Analysis

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RNA isolation and qRT-PCR was performed as previously described (38 ). Briefly, total RNA was extracted using Trizol reagent according to the manufacturer's protocols (Invitrogen). 1 μg of RNA was used for reverse transcription using a high capacity DNA reverse transcription kit (Applied Biosystems Cat No: 4368813), and qRT-PCR was performed using Power SYBR green PCR master mix (Applied Biosystems #4367659) according to manufacturer's protocols. The fold-change in expression was calculated by the 2–ΔΔCt methods. mRNA expression profiles were normalized to levels of housekeeping gene Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in each sample The primers used in qRT-PCR are listed in supplementary material and methods.
Kolapalli S.P., Sahu R., Chauhan N.R., Jena K.K., Mehto S., Das S.K., Jain A., Rout M., Dash R., Swain R.K., Lee D.Y., Rusten T.E., Chauhan S, & Chauhan S. (2020). RNA binding RING E3-ligase DZIP3/hRUL138 stabilizes Cyclin D1 to drive cell cycle and cancer progression. Cancer research, 81(2), 315-331.
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6

Quantitative Real-Time PCR Analysis

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The total RNA was extracted using Trizol reagent according to manufacturer’s protocols (Invitrogen). 2 μg of RNA was used for reverse transcription using high capacity DNA reverse transcription kit (Applied Biosystems Cat No: 4368813) and qRT-PCR was performed using Taqman master mix or Power SYBR green PCR master mix (Applied biosystems cat No: 4367659) according to manufacturer’s protocols on Step One Real-Time PCR system (Applied Biosystems). The following cycling program was used for SYBR green assay: initial denaturation at 95°C for 10 min; 40 cycles of 95°C for 15 s and 60°C for 60 s; followed by a melt curve step. For Taqman assay: Initial denaturation 50°C for 2 min and 95°C for 10 min; 40 cycles of 95°C for 15 s and 60°C for 60 s. mRNA expression profiles were normalized to levels of housekeeping gene Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) or β-Actin in each sample and the fold-change in expression was calculated by the 2−ΔΔCt method.
Mehto S., Jena K.K., Nath P., Chauhan S., Kolapalli S.P., Das S.K., Sahoo P.K., Jain A., Taylor G.A, & Chauhan S. (2019). The Crohn’s Disease Risk Factor IRGM Limits NLRP3 Inflammasome Activation by Impeding Its Assembly and by Mediating Its Selective Autophagy. Molecular Cell, 73(3), 429-445.e7.
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7

Quantitative RNA Analysis via qRT-PCR

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RNA isolation and qRT–PCR are performed exactly as described previously (Chauhan & Boyd, 2012). Briefly, total RNA was extracted using Trizol reagent according to manufacturer's protocols (Invitrogen). 2 μg of RNA was used for reverse transcription using high‐capacity DNA reverse transcription kit (Applied Biosystems #4368813), and qRT–PCR was performed using Power SYBR green PCR master mix (Applied Biosystems #4367659) according to manufacturer's protocols.
Jena K.K., Kolapalli S.P., Mehto S., Nath P., Das B., Sahoo P.K., Ahad A., Syed G.H., Raghav S.K., Senapati S., Chauhan S, & Chauhan S. (2018). TRIM16 controls assembly and degradation of protein aggregates by modulating the p62‐NRF2 axis and autophagy. The EMBO Journal, 37(18), e98358.
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8

RNA Extraction and qRT-PCR Analysis

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RNA isolation and qRT–PCR are performed exactly as described previously (Chauhan & Boyd, 2012 (link)). Briefly, total RNA was extracted using Trizol reagent according to manufacturer’s protocols (Invitrogen). 2 μg of RNA was used for reverse transcription using high-capacity DNA reverse transcription kit (Applied Biosystems #4368813), and qRT–PCR was performed using Power SYBR green PCR master mix (Applied Biosystems #4367659) according to manufacturer’s protocols.
Jena K.K., Kolapalli S.P., Mehto S., Nath P., Das B., Sahoo P.K., Ahad A., Syed G.H., Raghav S.K., Senapati S., Chauhan S, & Chauhan S. (2018). TRIM16 controls assembly and degradation of protein aggregates by modulating the p62‐NRF2 axis and autophagy. The EMBO Journal, 37(18), e98358.
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9

RNA Expression Analysis Using RT-qPCR

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RNA was extracted using TRIzol by following the manufacturer's protocol. The cDNA was synthesized using the high capacity DNA reverse transcription kit (Applied Biosystems, #4368813) and qRT–PCR was performed using TaqMan master mix (Applied Biosystems, #4369016) or Power SYBR green PCR master mix (Applied Biosystems, #4367659) according to manufacturer's protocol. For normalization of the assay, the housekeeping gene GAPDH or β‐Actin was used. The fold change in expression was calculated by the 2−ΔΔCt method.
Jena K.K., Mehto S., Nath P., Chauhan N.R., Sahu R., Dhar K., Das S.K., Kolapalli S.P., Murmu K.C., Jain A., Krishna S., Sahoo B.S., Chattopadhyay S., Rusten T.E., Prasad P., Chauhan S, & Chauhan S. (2020). Autoimmunity gene IRGM suppresses cGAS‐STING and RIG‐I‐MAVS signaling to control interferon response. EMBO Reports, 21(9), e50051.
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10

Quantitative Real-Time PCR Analysis of LOXL1 mRNA

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The generation of first-strand cDNA was performed using a high capacity DNA reverse transcription kit (Applied Biosystems) according to the manufacturer’s instructions, and the reverse transcriptase reaction products were used for quantitative real-time PCR, which was run in an AB7500 (Applied Biosystems) using TaqMan predesigned probes (Hs00935937_m1).
The amount of mRNA for LOXL1 in each sample was normalised to the amount of mRNA of the beta 2-tubulin reference gene in the same sample. Relative mRNA expression levels of all examined genes were measured using the comparative 2 ΔΔCT Ct method. Each plate contained three positive controls (samples previously confirmed by direct sequencing as both heterozygous and homozygous) and two negative controls.
Giardina E., Oddone F., Lepre T., Centofanti M., Peconi C., Tanga L., Quaranta L., Frezzotti P., Novelli G, & Manni G. (2014). Common sequence variants in the LOXL1 gene in pigment dispersion syndrome and pigmentary glaucoma. BMC Ophthalmology, 14, 52.
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