Peqgold rna kit

Manufactured by Avantor

Sourced in Germany

The PeqGold RNA kit is a laboratory equipment product designed for the isolation and purification of RNA from various biological samples. It provides a reliable and efficient method for extracting high-quality RNA for downstream applications.

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Lab products found in correlation

Script cdna synthesis kit, by Jena Biosciences (2 mentions) Sybr green mix, by Thermo Fisher Scientific (1 mentions) 7900ht thermal cycler, by Thermo Fisher Scientific (1 mentions) Mastercycler, by Eppendorf (1 mentions) Lightcycler 480 2, by Roche (1 mentions) Quantitect primer, by Qiagen (1 mentions) Verso cdna synthesis kit, by Thermo Fisher Scientific (1 mentions) Cfx96 real time pcr detection system, by Bio-Rad (1 mentions) Revertaid h minus first strand cdna synthesis kit, by Thermo Fisher Scientific (1 mentions) Sybr green pcr master mix, by Thermo Fisher Scientific (1 mentions) Quantstudio 7 flex real time pcr system, by Thermo Fisher Scientific (1 mentions) Quantitect primer assay, by Qiagen (1 mentions) Hiseq 4000, by Illumina (1 mentions) Nanodrop onec, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

PeqGOLD Total RNA Kit (181 citations) PeqGold Total RNA Isolation Kit (15 citations) Peqgold Total RNA Kit including DNase digestion (7 citations) PeqGOLD Bacterial RNA Kit (5 citations) PeqGOLD HP total RNA kit (4 citations) PeqGold RNA isolation kit (3 citations) PeqGold RNA extraction kits (2 citations) PeqGOLD Total RNA purification kit (2 citations) PeqGold Total RNA Micro Kit (2 citations) PeqGold total RNA extraction kit (2 citations)

7 protocols using peqgold rna kit

Hypoxia-Induced Changes in mRNA Expression

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Tumor cells were seeded at 10⁴ cells/well in 48 well plates in DMEM with 10% FBS. The cells were cultivated at either 20% or 2% oxygen (5% CO₂) for 48 h and harvested by passive lysis. RNA was isolated using the PeqGold RNA kit (Peqlab, Erlangen, Germany) according to the manufactures instructions.
mRNA-Expression was quantified by qRT-PCR on an 7900HT Thermal cycler (Applied Biosystems, Darmstadt, Germany) using SYBR green mix (Fermentas, Darmstadt, Germany). Sequences of primers used in qRT-PCR are given in supplemental table 1. All primer pairs were designed to span at least one intron to avoid amplification of contaminating gDNA. Expression levels of lox-family members were normalized against expression of rsp2960 (link).

Schütze F., Röhrig F., Vorlová S., Gätzner S., Kuhn A., Ergün S, & Henke E. (2015). Inhibition of Lysyl Oxidases Improves Drug Diffusion and Increases Efficacy of Cytotoxic Treatment in 3D Tumor Models. Scientific Reports, 5, 17576.

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RNA Extraction and qRT-PCR Analysis

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RNA extraction from cultured phLFs was performed using the PeqGold RNA Kit (Peqlab) according to the manufacturer’s instruction. The concentration of the isolated RNA was assessed spectrophotometrically at a wavelength of 260 nm (NanoDrop 1000). cDNA was synthesized with the GeneAMP Polymerase Chain Reaction (PCR) Kit (Applied Biosystems, Foster City, CA, USA) using random hexamers using 1 μg of isolated RNA for one 301 reaction. Denaturation was performed in an Eppendorf Mastercycler with the following settings: 302 303 lid = 45°C, 70°C for 10 min and 4°C for 5 min. Reverse transcription was performed in an Eppendorf Mastercycler with the following settings: lid = 105°C, 20°C for 10 min, 42°C for 60 min, and 99°C for 5 min. Quantitative reverse transcription PCR (qRT-PCR) reactions were performed in triplicates with SYBR Green I Master in a LightCycler 480II (Roche, Risch, Switzerland) with standard conditions: 95°C for 5 min followed by 45 cycles of 95°C for 5 s (denaturation), 59°C for 5 s (annealing), and 72°C for 20 s (elongation). Target genes were normalized to hypoxanthine-guanine phosphoribosyltransferase expression. All human primer sequences are documented in table S1.

Gerckens M., Schorpp K., Pelizza F., Wögrath M., Reichau K., Ma H., Dworsky A.M., Sengupta A., Stoleriu M.G., Heinzelmann K., Merl-Pham J., Irmler M., Alsafadi H.N., Trenkenschuh E., Sarnova L., Jirouskova M., Frieß W., Hauck S.M., Beckers J., Kneidinger N., Behr J., Hilgendorff A., Hadian K., Lindner M., Königshoff M., Eickelberg O., Gregor M., Plettenburg O., Yildirim A.Ö, & Burgstaller G. (2021). Phenotypic drug screening in a human fibrosis model identified a novel class of antifibrotic therapeutics. Science Advances, 7(52), eabb3673.

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RNA Extraction and Quantitative Real-Time PCR

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The peqGOLD RNA Kit (732-2868, Peqlab, Erlangen, Germany) was used to extract total RNA from tissues or cultured cells according to the manufacturer’s protocol. RNA was reverse transcribed into cDNA using the SCRIPT cDNA Synthesis Kit (PCR-5112, Jena Bioscience, Jena, Germany). Quantitative real-time PCR was performed with specific QuantiTect Primer assays (Qiagen, Hilden, Germany). HPRT or ACTB were used for normalization. Experiments were performed at least two times and representative data are shown.

Yu Y.Q., Herrmann A., Thonn V., Cordsmeier A., Neurath M.F., Ensser A, & Becker C. (2022). SMYD2 Inhibition Downregulates TMPRSS2 and Decreases SARS-CoV-2 Infection in Human Intestinal and Airway Epithelial Cells. Cells, 11(8), 1262.

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RNA Isolation and qPCR Analysis

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RNA isolation was performed with the RNA extraction kit peq gold RNA kit (peqlab, Erlangen, Germany). RNA concentrations were determined by measuring the absorbance at 260 and 280 nm. 1 µg of RNA was used for cDNA synthesis with the Verso cDNA Synthesis Kit (Thermo Fisher Scientific, Darmstadt, Germany). qPCR was performed by CFX96 Real-Time PCR Detection System (Bio-Rad Laboratories GmbH, Hercules, CA, USA). Gapdh and Actb served as internal housekeeping genes. Primer sequences are shown in Supplementary Table 1.

Kress J.M., Dio L.D., Heck L., Pulliero A., Izzotti A., Laarmann K., Fritz G, & Kaina B. (2019). Human primary endothelial cells are impaired in nucleotide excision repair and sensitive to benzo[a]pyrene compared with smooth muscle cells and pericytes. Scientific Reports, 9, 13800.

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Quantifying Gene Expression via qRT-PCR

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To quantify gene expression the indicated cells were subjected to isolation of total RNA using the PeqGold RNA kit (Peqlab, Erlangen, Germany), according to the manufacturer’s instructions. Between 500–1000 ng of RNA per sample were used for cDNA synthesis using the RevertAID H Minus First Strand cDNA synthesis kit (Thermo Fisher Scientific). Quantification of gene expression was completed using SYBR green-based qRT-PCR employing the SYBR™ Green PCR Master Mix (Thermo Fisher Scientific) and the QuantStudio 7 Flex Real-Time PCR System (Applied Biosystems, Thermo Fisher Scientific. Relative RNA levels of the genes of interest were calculated relative to GAPDH as the housekeeping gene, using the 2^−ΔΔCT-method [42 (link)]. At least three independent experiments in technical duplicates or triplicates were performed. All primers were purchased at Eurofins (Hamburg, Germany). Primer sequences are available upon request.

Wolf C., Smith S, & van Wijk S.J. (2022). Zafirlukast Induces VHL- and HIF-2α-Dependent Oxidative Cell Death in 786-O Clear Cell Renal Carcinoma Cells. International Journal of Molecular Sciences, 23(7), 3567.

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Quantitative Real-Time PCR Analysis

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Total RNA was extracted from tissues or cultured cells using peqGOLD RNA Kit (732-2868, Peqlab) according to the manufacturer’s protocol and reverse transcribed into cDNA using the SCRIPT cDNA Synthesis Kit (PCR-511L, Jena Bioscience). Quantitative real-time PCR was carried out using specific QuantiTect Primer assays (Qiagen). The mRNA expression of the reference gene hypoxanthine guanine phosphoribosyl transferase (HPRT) was used to normalize cDNA levels.

Yu Y.Q., Thonn V., Patankar J.V., Thoma O.M., Waldner M., Zielinska M., Bao L.L., Gonzalez-Acera M., Wallmüller S., Engel F.B., Stürzl M., Neurath M.F., Liebing E, & Becker C. (2022). SMYD2 targets RIPK1 and restricts TNF-induced apoptosis and necroptosis to support colon tumor growth. Cell Death & Disease, 13(1), 52.

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Transcriptome sequencing of iron-treated plants

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Total RNA was extracted with peqGOLD RNA kit (Peqlab; Erlangen, Germany) following the manufacturers' instructions. The genomic DNA was removed with RNAase-free with DNAase. The integrity of RNA was tested on a bleach agarose gel (Aranda et al., 2012) and for purity using NanoDrop OneC (Thermo Fisher Scientific, Braunschweig, Germany).
For transcriptomic analysis, approximately 10 µg of total RNA was used for high-throughput cDNA sequencing by Illumina HiSeq TM 4000 technology (BGI Genomics Co., Ltd -Denmark). RNAs derived from three biological replicates were used to generate the libraries. Eighteen libraries were constructed, from samples harvested seven days after the onset of treatments (control and +Fe). The cDNA libraries were prepared according to Illumina's protocols. After sequencing, read quality was checked and all low-quality reads (PHRED value < 20) were removed.

Wairich A., Oliveira B.H.N.d., Wu L., Murugaiyan V., Margis‐Pinheiro M., Fett J.P., Ricachenevsky F.K., & Frei M. (2020). Introgression fromOryza meridionalisinto domesticated riceOryza sativaresults in shoot-based iron tolerance.

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