Rocket script rt premix kit

Manufactured by Bioneer

The Rocket Script RT PreMix kit is a reagent designed for reverse transcription of RNA to cDNA. It contains all the necessary components, including reverse transcriptase enzyme, primers, and dNTPs, pre-mixed for convenience.

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Lab products found in correlation

Nanodrop spectrophotometer, by Eppendorf (2 mentions) Random hexamer primers, by Bioneer (2 mentions) Dnase 1, by Vivantis Technologies (2 mentions) Nanodrop spectrophotometry, by Eppendorf (1 mentions) Nanodrop nd 1000 spectrophotometer, by Thermo Fisher Scientific (1 mentions)

4 protocols using rocket script rt premix kit

RNA Extraction and cDNA Synthesis from Cardiac Muscle

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Total RNA was isolated from 100 mg of cardiac
muscles using RNX ^TM isolation reagent according to
the manufacturer’s procedure (CinaClon, Iran). Possible
DNA contamination was removed by treatment of
RNA (1 µg) with DNase I (2 U/µl) for 1 hour at 37oC
(Vivantis, Malaysia). Concentration of extracted RNA
was calculated at the wavelength of 260 nm using
NanoDrop spectrophotometer (Eppendorf, Germany). To
detect the purity of RNA, its optical density (OD) ratio at
260/280 nm was determined and samples having a ratio
>1.8 were used for cDNA synthesis. Reverse transcription
was carried out using the RocketScript RT PreMix kit
using 1 µg of RNA and random hexamer primers based
on manufacturer’s protocol (Bioneer Corporation, South
Korea). Reverse transcription was carried out at 42°C for
90 minutes followed by incubation at 70°C for 5 minutes.
cDNAs were stored at -20°C until used for real-time
polymerase chain reaction (PCR).

Ranjbar Kohan N., Nazifi S., Tabandeh M.R, & Lari M.A. (2018). Effect of L-Carnitine Supplementation on Apelin and Apelin Receptor (Apj) Expression in Cardiac Muscle of Obese Diabetic Rats. Cell Journal (Yakhteh), 20(3), 427-434.

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Pancreatic Gene Expression Analysis

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At the end of the experiment, animals were scarified and
pancreas tissues were immediately collected and frozen
at -70°C. Total RNA was isolated using RNX TM reagent
according to the manufacturer’s procedure (CinnaGen,
Iran). Concentration of extracted RNA was calculated at a
wavelength of 260 nm using nano drop spectrophotometry
(Eppendorf, Germany). To detect the purity of RNA, its
optical density (OD) ratio at 260/280 nm was determined and
samples with a ratio of >1.8 were used for cDNA synthesis.
Reverse transcription was carried out using the Rocket Script
RT PreMix kit using 1 µg of RNA and random hexamer,
based on manufacturer’s protocol (Bioneer Corporation,
South Korea). Reverse transcription was carried out at 42°C
for 90 minutes followed by incubation at 80°C for 3 minutes.
cDNAs were stored at -20°C until used in the real-time
polymerase chain reaction (PCR).

Erfani Majd N., Tabandeh M.R., Shahriari A, & Soleimani Z. (2018). Okra (Abelmoscus esculentus) Improved Islets Structure, and Down-Regulated PPARs Gene Expression in Pancreas of High-Fat Diet and Streptozotocin-Induced Diabetic Rats. Cell Journal (Yakhteh), 20(1), 31-40.

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RNA Extraction and cDNA Synthesis Protocol

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Total RNA was isolated from 100 mg of AT using RNX TM isolation reagent according to the manufacturer's procedure (Cina Clon, Iran). Possible DNA contamination was removed by the treatment of RNA (1 μg) with DNase I (2 U/μl) for 1 hr at 37°C (Vivantis, Malaysia). The concentration of extracted RNA was calculated at the wavelength of 260 nm using NanoDrop spectrophotometer (Eppendorf, Germany). For detecting the purity of the RNA, the optical density (OD) ratio at 260/280 nm was determined, and samples with a ratio >1.8 were used for cDNA synthesis. Reverse transcription was carried out using the Rocket Script RT PreMix kit with 1 μg of RNA and random hexamer primers based on the manufacturer's protocol (Bioneer Corporation, South Korea). Reverse transcription was carried out at 42°C for 90 min, followed by incubation at 70°C for 5 min. cDNAs were stored at −20°C to be used later for real‐time polymerase chain reaction (PCR).

Ranjbar Kohan N., Tabandeh M.R., Nazifi S, & Soleimani Z. (2020). L‐carnitine improves metabolic disorders and regulates apelin and apelin receptor genes expression in adipose tissue in diabetic rats. Physiological Reports, 8(23), e14641.

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Catharanthus Meristematic Cell Transcriptome

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Catharanthus meristematic cells were collected from 10-days-, 30-days-, and 50-day-old in vitro grown cultures. Four tissue samples were collected from each one of the CMCs and DDCs, which were maintained on their respective media. Total RNA was isolated using the RBC plant RNA isolation kit following the manufacturer's instructions. The RNA concentration and purity were measured with a NanoDrop ND-1000 spectrophotometer (Nanodrop Technologies, USA). The RNA integrity was checked by formaldehyde gel electrophoresis. Six hundred nanograms of total RNA was used for reverse-transcription with a Rocket script RT premix kit (Bioneer, South Korea). First-strand cDNA synthesis was carried out in a final volume of a 20-μL reaction mixture using oligo-dT (20 mer) primer according to the manufacturer's protocol. cDNA synthesis was carried out at 30 °C for 1 min, at 60 °C for 1 h, and at 95 °C for 5 min. The cDNA was diluted twice with nuclease-free water and stored at -20 °C until the quantitative reverse-transcription PCR (qRT-PCR) experiment.

Moon S.H., Venkatesh J., Yu J.W, & Park S.W. (2015). Differential induction of meristematic stem cells of Catharanthus roseus and their characterization. Comptes rendus biologies, 338(11).

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