Cdnasynthesis kit system

Manufactured by Roche

The CDNASynthesis Kit System is a laboratory equipment product designed for the synthesis of complementary DNA (cDNA) from RNA samples. The system provides the necessary components and reagents to perform the reverse transcription process, which is a fundamental step in various molecular biology applications such as gene expression analysis and cDNA library construction.

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Lab products found in correlation

Rna 6000 nano chip kit, by Agilent Technologies (2 mentions) Agilent 2100 bioanalyzer, by Agilent Technologies (2 mentions) Ms200 scanner, by Roche (2 mentions) Rneasy plus mini kit, by Qiagen (1 mentions) Deva 1, by Roche (1 mentions) Nucleospin rna kit, by Macherey-Nagel (1 mentions) Genelute pcr clean up kit, by Merck Group (1 mentions)

2 protocols using cdnasynthesis kit system

RNA Isolation and Microarray Analysis Protocol

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Biopsies stored at −80°C in RNAlater were used for the isolation of total RNA with the
RNeasy Plus Mini kit (Qiagen) according to the manufacturer’s instructions. RNA was
quantified by absorbance at 260/280 nm, and quality was evaluated using an Agilent 2100
Bioanalyzer and the RNA 6000 NanoChip kit (Agilent Technologies, Santa Clara, California).
Double-stranded complementary DNA (ds-cDNA) was generated from 5 µg of RNA using the cDNA
Synthesis Kit System (Roche Applied Science, Indianapolis, Indiana) and purified with the
GenElute polymerase chain reaction (PCR) Clean-up Kit (Sigma-Aldrich Quimica, Toluca,
Mexico). Afterward, the ds-cDNA was labeled with Cy3 and hybridized in a Human Gene
Expression Array 12 × 135 k (Roche Applied Science). Finally, the microarrays were scanned
on an MS200 Scanner (Roche Applied Science), and the data obtained were processed using
the DEVA 1.2 software (Roche Applied Science). Fluorescence intensities were normalized
using the RMA algorithm, and all data obtained were subsequently analyzed using the CLC
Main WorkBench version 7.0.3 software (Qiagen). Intensities of sensitive versus resistant
samples were compared and the genes were considered differentially expressed when the
intensities showed a difference of >1.5 and a P value <.05.

Garcia-Chagollan M., Carranza-Torres I.E., Carranza-Rosales P., Guzmán-Delgado N.E., Ramírez-Montoya H., Martínez-Silva M.G., Mariscal-Ramirez I., Barrón-Gallardo C.A., Pereira-Suárez A.L., Aguilar-Lemarroy A, & Jave-Suárez L.F. (2018). Expression of NK Cell Surface Receptors in Breast Cancer Tissue as Predictors of Resistance to Antineoplastic Treatment. Technology in Cancer Research & Treatment, 17, 1533033818764499.

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Transcriptome Analysis of Cell Lines and Samples

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Total RNA from the different cell lines, cervical scrapes, and biopsies was extracted with the NucleoSpin RNA Kit (Macherey-Nagel GmbH & Co. KG, Düren, Germany) according to the manufacturer's instructions. RNA was quantified by absorbance at 260/280 nm. For microarray analysis, RNA quality was determined using an Agilent 2100 Bioanalyzer and the RNA 6000 NanoChip Kit (Agilent Technologies, Santa Clara, CA, USA). Double stranded complementary DNA (cDNA) was generated from 10µg of RNA using the cDNA Synthesis Kit System (Roche Applied Science) and purified with the GenEluteTM PCR Clean-Up Kit (Sigma-Aldrich Quimica, S. A. de R.L. de C.V.); thereafter, the cDNA was labeled with Cy3 and hybridized in a Human Gene Expression Array 12x135K (Roche Applied Science). Microarrays were scanned on the MS200 Scanner and the data obtained was processed using DEVA ver 1.2 software (Roche Applied Science). Fluorescence intensities were normalized using the RMA algorithm and the information was subsequently analyzed with CLC Main WorkBench ver. 7.0.3 software. Genes that were differentially expressed with a fold-change >1.5 and that exhibited a permutation p-value <0.05 were selected.

Alvarado-Ruiz L., Martinez-Silva M.G., Torres-Reyes L.A., Pina-Sanchez P., Ortiz-Lazareno P., Bravo-Cuellar A., Aguilar-Lemarroy A, & Jave-Suarez L.F. (2016). HOXA9 is Underexpressed in Cervical Cancer Cells and its Restoration Decreases Proliferation, Migration and Expression of Epithelial-to-Mesenchymal Transition Genes. Asian Pacific journal of cancer prevention : APJCP, 17(3).

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