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Episcript reverse transcriptase

Manufactured by Illumina
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Episcript Reverse Transcriptase is a lab equipment product offered by Illumina. It is a reverse transcriptase enzyme used to convert RNA into complementary DNA (cDNA) molecules. The core function of Episcript Reverse Transcriptase is to catalyze the synthesis of single-stranded cDNA from an RNA template.

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Lab products found in correlation

Zymoprep direct zol rna microprep kit, by Zymo Research (2 mentions) Trizol reagent, by Thermo Fisher Scientific (2 mentions) Abi prism 7500 detection system, by Thermo Fisher Scientific (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Power sybr green pcr master mix, by Thermo Fisher Scientific (1 mentions) Cfx real time pcr detection system, by Bio-Rad (1 mentions) Cfx connect real time system, by Bio-Rad (1 mentions) Gotaq dna polymerase, by Promega (1 mentions) Random hexamer primer, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Reverse transcriptase (6 citations) Episcript (2 citations)

5 protocols using episcript reverse transcriptase

1

Quantification of Catalase Expression in Flies

Cited 1 time
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Whole heads were dissected from five adult male flies, and RNA was extracted using Trizol, followed by Zymoprep Direct-zol RNA MicroPrep kit (Zymo Research, Zurich, Switzerland, Cat # R2050) with DNAase I treatment. cDNA was generated from 100 ng of RNA using Episcript Reverse Transcriptase (Epicentre, Petaluma, CA, USA) and random hexamer primers. RT-qPCR was conducted as previously described [44 (link)]. Catalase gene expression was normalized to the geometric mean of two reference genes (RpL32, eIF1a). Statistical significance was determined using Student’s t-test.
Escobedo S.E., Stanhope S.C., Dong Z, & Weake V.M. (2022). Aging and Light Stress Result in Overlapping and Unique Gene Expression Changes in Photoreceptors. Genes, 13(2), 264.
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2

Quantitative Gene Expression Analysis of Immune Cells

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Total RNA samples were extracted from peritoneal neutrophils and prostate Gr (+)- and Gr (–)-sorted cells using TRIzol (Thermo Fisher, Carlsbad, CA). RNA was subsequently purified using Direct-zol RNA miniprep kit (Zymo Res., Irvine, CA) according to manufacturer's instructions. Then, 0.5 μg of RNA was used as a template for reverse transcription using EpiScript™ Reverse Transcriptase (Epicentre, Madison, WI) with random hexamer primers (Fermentas, Thermo Fisher, Carlsbad, CA). qPCR was performed with power SYBR green PCR master mix (Applied Biosystems, Foster City, CA) in a ABI Prism 7500 detection system (Thermo Fisher, Carlsbad, CA). The expression of ACTB was chosen as housekeeping gene. Data analysis was based on the 2−ΔΔCt method for normalization of raw data. All primers used are described in the Supplementary Table 1.
Scalerandi M.V., Peinetti N., Leimgruber C., Cuello Rubio M.M., Nicola J.P., Menezes G.B., Maldonado C.A, & Quintar A.A. (2018). Inefficient N2-Like Neutrophils Are Promoted by Androgens During Infection. Frontiers in Immunology, 9, 1980.
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3

RNA Extraction and RT-qPCR Analysis

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Total RNA was isolated using TRIzol® reagent (Invitrogen, Carlsbad, CA, USA). cDNA was made using EpiScript reverse transcriptase (Epicentre, WI, USA). Expression levels were measured by real-time PCR using SYBR and CFX real-time PCR detection system (Bio-Rad, Hercules, CA, USA). The PCR protocol was: 95 °C, 10 min, followed by 95 °C, 15 s and 60 °C, 30 s for 40 cycles. Rpl35a served as an internal control. Table 1 shows primer sequences.
Lin Y.T., Yu Z., Tsai S.C., Hsu P.H, & Chen J.C. (2020). Neuropeptide FF receptor 2 inhibits capsaicin-induced CGRP Upregulation in mouse trigeminal ganglion. The Journal of Headache and Pain, 21(1), 87.
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4

Quantitative RT-PCR for Drosophila Gene Expression

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RNA was isolated using the Zymoprep Direct-zol RNA MicroPrep kit (Zymo Research) and treated with DNAse I as per the kit protocol. cDNA was generated from 250 ng of RNA using Episcript Reverse Transcriptase (Epicentre) using either oligo dTs or random hexamer primers as indicated. qPCR was conducted using Evagreen 2X Mix (Biotium) and the CFX Connect Real-time system (Biorad). Quantities were determined relative to a 4-fold dilution series of wild-type (OregonR) cDNA. Primers against SAGA-regulated genes were taken from previous studies [40 (link)]. New primers used in this study are as follows: SF3B5 5′-GCAAAATGGGTGAACGCTAC-3′ and 5′-AGCCACTCGAACTTTGTGGT-3′, Sas10 5′-ACCGGTGCTCAACTACGTTC-3′ and 5′-GCTCCTCGATCAGATCCTTG-3′, Oda (unspliced) 5’-CCGTGCAAAAAGTGAATGTG-3’ and 5’-GCCAACCTGGAGAACGTCTA -3’, Sap47 (unspliced) 5’-ATCGATATTCCGCTTGTTGC-3’ and 5’-GCGCAAGTTTGATATTGTCG-3’, exba (unspliced) 5’-GAGCCCAAGGACAGGATTG-3’ and 5’-TGCTTGAACGTCTGGAACAG-3’, Crc (unspliced) 5’-′CGGACGAGTTGTCAACAGAA-3’ and 5’-′TCTGAAGATGCACCGAATTG-3’.
Stegeman R., Spreacker P.J., Swanson S.K., Stephenson R., Florens L., Washburn M.P, & Weake V.M. (2016). The spliceosomal protein SF3B5 is a novel component of Drosophila SAGA that functions in gene expression independent of splicing. Journal of molecular biology, 428(18), 3632-3649.
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5

Total RNA Extraction and RT-PCR Analysis

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Total RNA was extracted with TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer's instructions. Thereafter, 1 μg of total RNA was used as a template for reverse transcription using EpiScript™ Reverse Transcriptase (Epicentre, Madison, WI) with random hexamer primers (Thermo Fisher Scientific, Waltham, MA USA). The cDNA obtained was subjected to PCR amplification using GoTaq DNA Polymerase (Promega, Madison, WI), and following this, PCR protocols were performed according to the manufacturer's instructions. PCR amplification was performed in 10 μl using a specific primers sets (Table 1), and the PCR amplified products were visualized by electrophoresis onto 2% agarose gels by GelSampleRed TM (Biotium) staining, whit the band densitometric analysis being carried out using ImageJ software.
Sabatino M.E., Grondona E., Sosa L.D.V., Mongi Bragato B., Carreño L., Juarez V., da Silva R.A., Remor A., de Bortoli L., de Paula Martins R., Pérez P.A., Petiti J.P., Gutiérrez S., Torres A.I., Latini A, & De Paul A.L. (2018). Oxidative stress and mitochondrial adaptive shift during pituitary tumoral growth. Free radical biology & medicine, 120.
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