Picopure rna isolation kit

Manufactured by Qiagen

Sourced in Australia

The PicoPure RNA Isolation Kit is a laboratory tool designed to extract and purify RNA from small samples. It is a compact and efficient kit that can be used to isolate RNA from a variety of sources, including tissue, cells, and bodily fluids.

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Lab products found in correlation

Rnase free dnase set, by Qiagen (3 mentions) Ovation ultralow library system dr multiplex system, by Tecan (2 mentions) Ovation rna seq system v2, by Tecan (2 mentions) Reverse transcription kit with rnase inhibitor, by Thermo Fisher Scientific (1 mentions) Rneasy mini micro kit, by Qiagen (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Gotaq qpcr master mix, by Promega (1 mentions) Nextseq 500 instrument, by Illumina (1 mentions) Nextseq 500 high output reagent kit, by Illumina (1 mentions) High sensitivity rna screen tape system, by Agilent Technologies (1 mentions) Smart seq v4 ultra low input rna kit, by Takara Bio (1 mentions) Rneasy plus mini kit, by Qiagen (1 mentions) Nextera xt dna sample preparation kit, by Illumina (1 mentions) Picopure rna isolation kit, by Arcturus (1 mentions) Nd 1000, by Avantor (1 mentions) First strand cdna synthesis kit, by Thermo Fisher Scientific (1 mentions) Lightcycler 96, by Roche (1 mentions) Dnase treatment, by Qiagen (1 mentions) Pixcell 2 lcm system, by Arcturus (1 mentions) Dnase 1 digest, by Qiagen (1 mentions)

Close spelling variants of this product

RNA isolation kit

9 protocols using picopure rna isolation kit

RNA Extraction and RT-qPCR Analysis

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Total cellular RNA was extracted by Trizol (Invitrogen) phase separation followed by purification using RNeasy Mini/Micro kit (Qiagen), or by using the Arcturus PicoPure RNA Isolation Kit. Reverse transcription was carried out using high-capacity cDNA Reverse Transcription Kit with RNase Inhibitor (Applied Biosystems), or with SuperScript III First Strand Synthesis System for RT-polymerase chain reaction (PCR) (Invitrogen). Real-time PCR was carried out with primers listed in the Supporting Information using GoTaq qPCR Master Mix (Promega).

Fric J., Lim C.X., Mertes A., Lee B.T., Viganò E., Chen J., Zolezzi F., Poidinger M., Larbi A., Strobl H., Zelante T, & Ricciardi-Castagnoli P. (2014). Calcium and Calcineurin-NFAT Signaling Regulate Granulocyte-Monocyte Progenitor Cell Cycle via Flt3-L. Stem Cells (Dayton, Ohio), 32(12), 3232-3244.

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Transcriptome Profiling of Oocytes

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Total RNA was extracted from 25 wild-type or Sirena1 knock-out oocytes from 11 (GV) or 13–16 (MII) week old animals using the PicoPure RNA Isolation Kit with on-column genomic DNA digestion according to the manufacturer's instructions (Qiagen). Each sample was spiked with 0.2 pg synthesized Renilla luciferase mRNA before extraction. RNA-Seq libraries were constructed using the Ovation RNA-Seq system V2 (NuGEN) followed by Ovation Ultralow Library system (DR Multiplex System, NuGEN). RNA-Seq libraries were pooled and sequenced using 65-nt single-end-sequencing using Illumina HiSeq. All high-throughput datasets used in this work are listed in Supplementary Table S2. The code used for sequencing data analysis is available through GitHub: https://github.com/fhorvat/bioinfo_repo/tree/master/papers/Sirena1_2019.

Ganesh S., Horvat F., Drutovic D., Efenberkova M., Pinkas D., Jindrova A., Pasulka J., Iyyappan R., Malik R., Susor A., Vlahovicek K., Solc P, & Svoboda P. (2020). The most abundant maternal lncRNA Sirena1 acts post-transcriptionally and impacts mitochondrial distribution. Nucleic Acids Research, 48(6), 3211-3227.

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Comparative Oocyte Transcriptome Analysis

Cited 3 times

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Total RNA was extracted from 25 wild-type or D6Ertd527e1^–/– fully-grown oocytes from 8–10-week-old animals using PicoPure RNA Isolation Kit with on-column genomic DNA digestion according to the manufacturer’s instruction (Qiagen). RNA-Seq libraries were constructed using the Ovation RNA-Seq system V2 (NuGEN) followed by Ovation Ultralow Library system (DR Multiplex System, NuGEN). RNA-Seq libraries were pooled and sequenced using 65-nt single-end-sequencing using Illumina HiSeq. D6Ertd527e1^−/− oocyte sequencing data were deposited in GEO (https://www.ncbi.nlm.nih.gov/geo/) as GSE213820. Cricetulus griseus oocyte sequencing data were also deposited under GSE213820. Remaining RNA-seq data were published previously and were obtained from GEO database: Mus musculus data accession: GSE116771 [27 (link)], Mesocricetus auratus data accession: GSE116771 [12 (link)] and GSE169528 [33 (link)], Rattus norvegicus data accession: GSE137562 [35 (link)], and Acomys cahirinus data accession: PRJNA436818 [36 ].

Petrzilek J., Pasulka J., Malik R., Horvat F., Kataruka S., Fulka H, & Svoboda P. (2022). De novo emergence, existence, and demise of a protein-coding gene in murids. BMC Biology, 20, 272.

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Neuron-specific Transcriptome Analysis

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Another set of frozen postmortem brain samples from the same brain bank was used for a neurotranscriptome study as described previously [15 (link), 16 (link)]. Briefly, brain sections were stained with 1% neutral red, and pyramidal neurons were identified by their characteristic size, shape, and location. Total RNA was isolated from the cell lysate with the Arcturus PicoPure RNA Isolation Kit with DNase I treatment using Qiagen’s RNase-free DNase Set. Isolated total RNA from each sample of ~ 500 neurons was double-round amplified, cleaned, and biotin labeled with Affymetrix’s GeneChip Two-Cycle Target Labeling kit with a T7 promoter and Ambion’s MEGAscript T7 High Yield Transcription Kit per the manufacturer’s protocol. Amplified and labeled cRNA was quantitated on a spectrophotometer and run on a 1% Tris-acetate-EDTA gel to check for an evenly distributed range of transcript sizes.

Yin J., Han P., Song M., Nielsen M., Beach T.G., Serrano G.E., Liang W.S., Caselli R.J, & Shi J. (2018). Amyloid-β Increases Tau by Mediating Sirtuin 3 in Alzheimer’s Disease. Molecular neurobiology, 55(11), 8592-8601.

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RNA-Seq Library Preparation from Sorted Cells

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Cells were sorted as described in “Tissue Preparation and Flow Cytometry” and immediately lysed with RLT Buffer from the Qiagen RNeasy Plus Mini kit or RNA extraction buffer from PicoPure RNA isolation kit (Arcturus Bioscience, Inc.). Cell lysates were stored at −80°C until RNA was extracted. RNA isolations were performed using the Qiagen RNeasy Plus Mini kit or PicoPure RNA isolation kit. RNA quality and quantity were measured using the Agilent Technologies High Sensitivity RNA ScreenTape System. SMART-Seq v4 Ultra Low Input RNA kit (Clontech Laboratories, Inc.) was used for full-length cDNA synthesis, and the Nextera XT DNA sample preparation kit (Illumina Inc.) was used for library preparation. DNA libraries were sequenced on an Illumina NextSeq 500 instrument with a target read depth of ∼10 million aligned reads per sample. The pool was denatured and diluted, resulting in a 2.5 pM DNA solution. PhiX control was spiked at 1%, and the pool was sequenced by 1 × 75 cycles using the NextSeq 500 High Output reagent kit (Illumina Inc.).

Tsai F., Homan P.J., Agrawal H., Misharin A.V., Abdala-Valencia H., Haines GK I.I.I., Dominguez S., Bloomfield C.L., Saber R., Chang A., Mohan C., Hutcheson J., Davidson A., Budinger G.R., Bouillet P., Dorfleutner A., Stehlik C., Winter D.R., Cuda C.M, & Perlman H. (2017). Bim suppresses the development of SLE by limiting myeloid inflammatory responses. The Journal of Experimental Medicine, 214(12), 3753-3773.

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Quantifying BDNF mRNA Expression in Mice

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RNA purification was performed using the Arcturus PicoPure RNA Isolation Kit (#KIT0202, KIT0204), including digestion of genomic DNA using a RNase free DNase set (QIAGEN, #79254). RNA concentration was measured with a Nanodrop Spectrophotometer (Peqlab; ND-1000, RRID:SCR_016517); 20 ng purified RNA was used for reverse transcriptase-mediated cDNA synthesis, using the Thermo Fisher Scientific First Strand cDNA Synthesis Kit (#K1612). cDNA was diluted 1:5 in RNase free H₂O, and 2 µl cDNA was used for qRT-PCR using a Roche Diagnostics LightCycler 96 with the following primers against total BDNF (forward: 5'-AAATTACCTGGATGCCGCAAAC-3'; reverse: 5'-CGCTGTGACCCACTCGCTAA-3') and mouse GAPDH (forward: 5'-GCAAATTCAACGGCACA-3'; reverse: 5'-CACCAGTAGACTCCACGAC-3'). Results were exported to Microsoft Excel and analyzed with GraphPad Prism 6.0 for statistical analysis. BDNF values were normalized to GAPDH, and BDNF mRNA levels were calculated in percent of mean BDNF mRNA levels in P21 sedentary mice.

Andreska T., Rauskolb S., Schukraft N., Lüningschrör P., Sasi M., Signoret-Genest J., Behringer M., Blum R., Sauer M., Tovote P, & Sendtner M. (2020). Induction of BDNF Expression in Layer II/III and Layer V Neurons of the Motor Cortex Is Essential for Motor Learning. The Journal of Neuroscience, 40(33), 6289-6308.

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Microdissection of Colorectal Cancer Tissues

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For microdissection, colon cancer tissues and paired noncancerous colon tissues were obtained from patients undergoing surgical resection. The sample from the tumor tissue was mounted in Tissue-Tek OCT compound (Sakura Finechemicals, Tokyo, Japan) and frozen. Each sample was then cut into 10–20 serial sections with a thickness of 10 μm, and sections were mounted on uncoated glass slides. The parts containing areas of cancer cells were identified using hematoxylin–eosin staining and then microdissected according to the standard laser capture procedure^{32 (link), 33 (link)} using a PixCell II LCM system (Arcturus Engineering, Mountain View, CA, USA/Olympus, Tokyo, Japan). Total RNA was extracted from Laser-captured cell nests by using PicoPure RNA Isolation Kit according to the manufacturer's protocol, including on-column DNase treatment (Qiagen, Chadstone Centre, VIC, Australia).

Guo S.T., Chi M.N., Yang R.H., Guo X.Y., Zan L.K., Wang C.Y., Xi Y.F., Jin L., Croft A., Tseng H.Y., Yan X.G., Farrelly M., Wang F.H., Lai F., Wang J.F., Li Y.P., Ackland S., Scott R., Agoulnik I.U., Hondermarck H., Thorne R.F., Liu T., Zhang X.D, & Jiang C.C. (2015). INPP4B is an oncogenic regulator in human colon cancer. Oncogene, 35(23), 3049-3061.

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TCR and Transcriptome Analysis of Sorted TILs

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Sorted TILs were cultured in complete T cell medium supplied with 100 IU/mL rhIL2 for 24 hours. RNA samples were then extracted from 10000 cells of each population using Picopure™ RNA isolation kit (Qiagen RNase-Free DNase set were used to digest trace DNA). TCRβ sequencing was performed and analyzed by iRepertoire, Inc. RNA-seq was performed by Next Generation Sequencing Core at Scripps Research. The library for RNA-seq was prepared using SMARTseq HT kit and sequencing was performed using NextSeq500 sequencing platform. The reads were trimmed for the adapter sequences using cutadapt 1.18 with Python 3.6.3 and the trimmed reads were mapped to the reference genome using the STAR aligner 2.5.2a. Gene abundance was estimated with python 2.7.11, and HTSeq 0.11.0. PCA and the differential gene expression analyses between different cell populations was performed using DESeq2 package (Love et al., 2014 (link)) in R. Significantly different threshold was Benjamini–Hochberg FDR (p.adjust) <0.05 and |log₂(foldchange)| ≥0.6. Gene ontology over-representation analysis was performed using clusterProfiler package (Yu et al., 2012 (link)) in R. GSEA analysis was performed using GSEA 4.0.3 (Broad Institute).

Liu Z., Li J.P., Chen M., Wu M., Shi Y., Li W., Teijaro J.R, & Wu P. (2020). Detecting Tumor Antigen-specific T cells via Interaction Dependent Fucosyl-biotinylation. Cell, 183(4), 1117-1133.e19.

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Single-Cell RNA-Seq Library Prep

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Between 1000 and 5000 cells were FACS-sorted directly into the extraction buffer (PicoPure RNA Isolation Kit, Applied Biosystems), snap frozen, and stored at −80°C. RNA was extracted using the PicoPure RNA Isolation Kit including an on-column DNase I digest (Qiagen, 79254) following the manufacturer’s instructions. RNA-seq libraries were generated from 2 to 5 ng of total RNA using the NEBNext Single Cell/Low Input RNA Library Prep Kit for Illumina (NEB, E6420) and NEBNext Multiplex Oligos for Illumina (NEB, E7335S). Library quality was accessed by TapeStation 2200 (Agilent) with High Sensitivity D1000 ScreenTape (Agilent), quantified, and multiplexed. The multiplexed libraries were subjected to paired-end 100–base pair (bp) sequencing on HiSeq 4000 sequencing system (Illumina), resulting in >30 million reads per sample.

Gruhn W.H., Tang W.W., Dietmann S., Alves-Lopes J.P., Penfold C.A., Wong F.C., Ramakrishna N.B, & Surani M.A. (2023). Epigenetic resetting in the human germ line entails histone modification remodeling. Science Advances, 9(3), eade1257.

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