Total RNA from cell lines was extracted using TRI Reagent Solution (Thermo Fisher Scientific) or Direct-zol RNA Miniprep (Zymo Research) according to the manufacturer’s instructions. Total RNA from FFPE samples was extracted from 5 sections of 5 µm thickness using High Pure FFPET RNA isolation kit (Roche) according to the manufacturer’s instructions. Total RNA was purified from plasma using two extraction kits, according to the available volume of plasma samples. MiRNeasy Serum/Plasma Advanced Kit (QIAGEN) was used, according to the manufacturer’s instructions, to extract RNA from 600 µl of plasma eluted in a final volume of 15 µl, while Plasma/Serum Circulating and Exosomal RNA Purification Mini Kit (Norgen) was employed, according to the manufacturer’s instructions, to extract RNA from 1 ml of plasma eluted in a final volume of 100 µl. Only RNA samples extracted with the same method were pooled together for analysis.
Plasma serum circulating and exosomal rna purification mini kit
Manufactured by Norgen Biotek
Sourced in Canada
The Plasma/Serum Circulating and Exosomal RNA Purification Mini Kit is a laboratory equipment designed for the isolation and purification of RNA from plasma, serum, and exosomal samples. The kit utilizes a specialized protocol and proprietary reagents to efficiently extract high-quality RNA from these sample types.
Automatically generated - may contain errors
Lab products found in correlation
Tri reagent solution, by Thermo Fisher Scientific (2 mentions)
Mirneasy serum plasma advanced kit, by Qiagen (2 mentions)
Direct zol rna miniprep, by Zymo Research (2 mentions)
High pure ffpet rna isolation kit, by Roche (2 mentions)
Dsdna hs assay kit, by Thermo Fisher Scientific (1 mentions)
Hs ngs fragment kit, by Agilent Technologies (1 mentions)
Smarter stranded total rna seq kit v3 pico input mammalian, by Takara Bio (1 mentions)
Nextseq 2000, by Illumina (1 mentions)
Qubit 3.0 fluorometer, by Thermo Fisher Scientific (1 mentions)
Agilent 5200 fragment analyzer, by Agilent Technologies (1 mentions)
Nextseq 500 550, by Illumina (1 mentions)
Turbo dnase, by Thermo Fisher Scientific (1 mentions)
5 protocols using plasma serum circulating and exosomal rna purification mini kit
1
RNA Extraction from Cell Lines and FFPE Samples
2
Circulating miRNA Extraction from Prostate Cancer Samples
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Blood samples were obtained simultaneously for PSA measurement and serum miRNA extraction. Serum was collected within 2 h of blood draw. Mean and median time after radical prostatectomy to sample collection was 31 and 9 months, respectively (range = 3 weeks to 16 years). Circulating miRNAs were collected from serum using the Plasma/Serum Circulating and Exosomal RNA Purification Mini Kit (Norgen Biotek Corp., Canada).
3
Serum miRNA Profiling Protocol
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A fixed volume of serum (600 μl per sample) from each patient was used for miRNA profiling and quantitative real-time PCR detection. Total RNA was extracted with the Plasma/Serum Circulating and Exosomal RNA Purification Mini Kit (51000, NorgenBiotek Corporation, Canada), following the manufacturer’s instructions. Prior to RNA isolation from serum, a non-human miRNA, cel-miR-39, was spiked into the serum with a final concentration of 0.05 nM as a reference. At last, the extracted RNA dissolved in 100 μl nuclease-free water and eluted from spin-column was first dried by vacuum centrifugation at low temperature and then re-dissolved in RNA dephosphorylation reaction buffer for miRNA microarray analysis or nuclease-free water for qRT-PCR analysis.
4
RNA Extraction from Cell Lines and FFPE Samples
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Total RNA from cell lines was extracted using TRI Reagent Solution (Thermo Fisher Scientific) or Direct-zol RNA Miniprep (Zymo Research) according to the manufacturer’s instructions. Total RNA from FFPE samples was extracted from 5 sections of 5 µm thickness using High Pure FFPET RNA isolation kit (Roche) according to the manufacturer’s instructions. Total RNA was purified from plasma using two extraction kits, according to the available volume of plasma samples. MiRNeasy Serum/Plasma Advanced Kit (QIAGEN) was used, according to the manufacturer’s instructions, to extract RNA from 600 µl of plasma eluted in a final volume of 15 µl, while Plasma/Serum Circulating and Exosomal RNA Purification Mini Kit (Norgen) was employed, according to the manufacturer’s instructions, to extract RNA from 1 ml of plasma eluted in a final volume of 100 µl. Only RNA samples extracted with the same method were pooled together for analysis.
5
Plasma cfRNA Extraction and Library Prep
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Plasma samples were received on dry ice and stored at −80°C until processed. Prior to cfRNA extraction, plasma samples were thawed at room temperature and centrifuged at 1300 x g for 10 minutes at 4°C. cfRNA was extracted from plasma (300–1,000 μl) using the Norgen Plasma/Serum Circulating and Exosomal RNA Purification Mini Kit (Norgen, 51000). Extracted RNA was DNase treated with 14 μl of 10 μl DNase Turbo Buffer (Invitrogen, AM2238), 3 μl DNase Turbo (Invitrogen, AM2238), and 1 μl Baseline Zero DNase (Lucigen-Epicenter, DB0715K) for 30 minutes at 37°C, then concentrated into 12 μl using the Zymo RNA Clean and Concentrated Kit (Zymo, R1015).
Sequencing libraries were prepared from 8 μl of concentrated RNA using the Takara SMARTer® Stranded Total RNA-Seq Kit v3 - Pico Input Mammalian (Takara, 634485) and barcoded using the SMARTer® RNA Unique Dual Index Kit (Takara, 634451). Library concentration was quantified using the Qubit™ 3.0 Fluorometer (Invitrogen, Q33216) with the dsDNA HS Assay Kit (Invitrogen, Q32854). Libraries were quality-controlled using the Agilent Fragment Analyzer 5200 (Agilent, M5310AA) with the HS NGS Fragment Kit (Agilent, DNF-474–0500) and pooled to equal concentrations. Each pool was sequenced using both the Illumina NextSeq 500/550 platform (paired-end, 150 bp) and the Illumina NextSeq 2000 platform (paired-end, 100 bp).
Sequencing libraries were prepared from 8 μl of concentrated RNA using the Takara SMARTer® Stranded Total RNA-Seq Kit v3 - Pico Input Mammalian (Takara, 634485) and barcoded using the SMARTer® RNA Unique Dual Index Kit (Takara, 634451). Library concentration was quantified using the Qubit™ 3.0 Fluorometer (Invitrogen, Q33216) with the dsDNA HS Assay Kit (Invitrogen, Q32854). Libraries were quality-controlled using the Agilent Fragment Analyzer 5200 (Agilent, M5310AA) with the HS NGS Fragment Kit (Agilent, DNF-474–0500) and pooled to equal concentrations. Each pool was sequenced using both the Illumina NextSeq 500/550 platform (paired-end, 150 bp) and the Illumina NextSeq 2000 platform (paired-end, 100 bp).
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