The mirVana PARIS Kit (Ambion, Austin, TX) was used to extract total RNA from 200 µL serum or exosomes following the given protocol. For normalization, 5 µL of synthetic C. elegansmiR‐39 (5 nM/L, RiboBio, Guangzhou, China) was added to each sample after the addition of denaturing solution (Ambion, Austin, TX). Tissue samples were processed using Trizol (Invitrogen, Carlsbad, CA) to extract total RNA. Acquired total RNA was lysed in 100 µL RNase‐free water and restored at −80°C until use. The NanoDrop ND‐1000 spectrophotometer (NanoDrop, Wilmington, DE) was used to measure the concentration and purity of RNA.
Synthetic c elegans mir 39
Manufactured by RiboBio
Sourced in China
Synthetic C.elegans miR-39 is a laboratory reagent used as an internal control in small RNA quantification experiments. It is a synthetic version of the miR-39 microRNA found in the model organism Caenorhabditis elegans.
Automatically generated - may contain errors
Lab products found in correlation
Denaturing solution, by Thermo Fisher Scientific (5 mentions)
Trizol, by Thermo Fisher Scientific (4 mentions)
Mirvana paris kit, by Thermo Fisher Scientific (4 mentions)
Ultraviolet spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Rneasy mini kit, by Qiagen (1 mentions)
5 protocols using synthetic c elegans mir 39
1
Extraction and Quantification of Serum/Exosomal miRNA
2
Serum and Exosome RNA Extraction
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Total RNA was extracted from 200μl serum or exosome using the mirVana PARIS Kit (Ambion, Austin, TX, USA) according to the manufacturer’s protocol. 5μl of synthetic C.elegans miR-39 (5 nM/L, RiboBio, Guangzhou, China) was added to each sample after the addition of denaturing solution (Ambion, Austin, TX, USA) for normalization. Trizol (Invitrogen, Carlsbad, CA, USA) was used to extract total RNA from tissue samples. Finally, total RNA was dissolved in 100μl RNase-free water and kept at -80°C until further analysis. The ultraviolet spectrophotometer was applied to evaluate the concentration and purity of the extracted total RNA.
3
Total RNA Extraction from Serum and Tissue
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Total RNA was extracted from 200 μL serum or exosomes using the mirVana PARIS Kit (Ambion, Austin, TX, USA) following the given protocol. 5 μL of synthetic C.elegans miR-39 (5 mmol/L, RiboBio, Guangzhou, China) was added for sample-to-sample normalization after the addition of denaturing solution (Ambion, Austin, TX, USA). Total RNA of tissue samples was obtained using Trizol (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's protocol. After being extracted, total RNA was dissolved in 100 μL RNase-free water and stored at −80°C until analysis. The concentration and purification of the RNA sample were assessed by the ultraviolet spectrophotometer.
4
Plasma RNA Extraction with Normalization
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The total RNA from 400 μl plasma was extracted using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) and purified using an RNeasy Mini kit (Qiagen, Hilden, German) according to the manufacturer’s instructions. Briefly, 10 nM synthetic C. elegans miR-39 (RiboBio, China) was spiked-in to each sample after the addition of denaturing solution (Ambion, Austin, TX, USA) for normalization of the between-sample variation. RNA concentrations were determined by measuring the sample absorbance at 260 nm with the ultraviolet spectrophotometer (Thermo Fisher Scientific, Inc., Waltham, MA, USA).
5
RNA Extraction from Tissue, Serum, and Exosomes
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For tissue samples, Trizol (Invitrogen, Carlsbad, CA, USA) was used to isolate total RNA in consistent with the manufacturer's protocol. Total RNA from serum or exosomes was extracted with the mirVana PARIS Kit (Ambion, Austin, TX, USA) according to the instructions. After the addition of denaturing solution (Ambion, Austin, TX, USA), 5 μL synthetic C. elegans miR-39 (5 nmol/L, RiboBio, Guangzhou, China) was added into each sample for normalization of the sample-to-sample variation. Total RNA was lysed in 100 μL RNase-free water and then stored at -80 °C for further analysis.
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