Cell-free plasma was isolated via a two-step protocol (2500 rpm at room temperature for 10 min and 14000×g at 4℃ for 10 min) within 2 h after collection to prevent contamination of the cellular nucleic acids. The resulting plasma was transferred to new tubes and stored at -80℃. The plasma was transferred to RNase/DNase-free tubes and stored at -80℃ until RNA isolation. The total RNA was isolated from the plasma using a mirVana PARIS isolation kit (Ambion, Austin, TX, USA) according to the manufacturer's instructions for plasma samples without enrichment for small RNAs. Briefly, 400 µL of plasma was used to extract the total RNA. Each sample was eluted in 40 µL of RNAse-free water. Cancer and non-cancer tissues from LSCC cases were immediately flash frozen in liquid nitrogen, and stored at -80℃ until RNA isolation using a mirVana PARIS isolation kit (Ambion, Austin, TX, USA) according to the manufacturer's instructions, which were similar to those described above. The average levels of miR-155 expression in tissues and sera were normalized relative to the average amounts of U6 snRNAm, using the 2-ΔΔCT method. RNA quality was measured using a denaturing 15% polyacrylamide gel.
Mirvana paris isolation kit
Manufactured by Thermo Fisher Scientific
Sourced in United States, Lithuania
The MirVana PARIS isolation kit is a tool designed for the rapid and efficient isolation of total RNA, including small RNAs such as microRNAs, from a variety of sample types. The kit utilizes a proprietary method to isolate RNA while effectively removing DNA, proteins, and other contaminants.
Automatically generated - may contain errors
Lab products found in correlation
Bulge loop mirna qpcr primer set, by RiboBio (2 mentions)
2100 bioanalyzer, by Agilent Technologies (2 mentions)
Tempus spin rna isolation kit, by Thermo Fisher Scientific (1 mentions)
High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (1 mentions)
Qubit 4 fluorometer, by Thermo Fisher Scientific (1 mentions)
7900ht fast real time pcr system, by Thermo Fisher Scientific (1 mentions)
Nanodrop nd 1000, by Thermo Fisher Scientific (1 mentions)
Tb green premix ex taq, by Takara Bio (1 mentions)
7900ht fast rt pcr system, by Thermo Fisher Scientific (1 mentions)
Edta coated vacutainer tubes, by BD (1 mentions)
Universal cdna synthesis kit, by Qiagen (1 mentions)
C elegans mir 39, by Thermo Fisher Scientific (1 mentions)
Nanodrop spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Recoverall total nucleic acid isolation kit for ffpe, by Thermo Fisher Scientific (1 mentions)
Dnase 1 dna free kit, by Thermo Fisher Scientific (1 mentions)
23 protocols using mirvana paris isolation kit
1
Plasma RNA Isolation and miR-155 Quantification
2
Comprehensive mRNA and miRNA Isolation from Blood
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total mRNA isolation from whole-blood samples was performed with the use of a Tempus Spin RNA Isolation Kit (Ambion) in accordance with the manufacturer's instructions. Final RNA concentration and purity were measured with a Qubit 4 Fluorometer (Invitrogen). A 1-μg portion of total mRNA per sample was reverse-transcribed with a High Capacity cDNA Reverse Transcription Kit (Invitrogen) in accordance with the manufacturer's instructions. Incubation was at 25°C for 10 min, reverse transcription was at 37°C for 120 min, and inactivation was at 85°C for 5 min. cDNA containing 180 ng RNA/sample was isolated from blood lymphocytes of CD or PD participants in all time periods.
Total RNA (including miRNA) was isolated from plasma samples with the use of a mirVana PARIS Isolation Kit (Applied Biosystems) in accordance with the manufacturer's protocol as described elsewhere (22 ). We selected 7 human circulating miRNAs (hsa-miR-15a-5p, hsa-miR-21-5p, hsa-miR-29b-3p, hsa-miR-126-3p, hsa-miR-192-5p, hsa-miR-223-3p, and hsa-miR-375) widely related to glucose metabolism, IR status, pre-diabetic status and biomarkers of T2D, based on the use of updated reviews and databases (22 ).
Total RNA (including miRNA) was isolated from plasma samples with the use of a mirVana PARIS Isolation Kit (Applied Biosystems) in accordance with the manufacturer's protocol as described elsewhere (22 ). We selected 7 human circulating miRNAs (hsa-miR-15a-5p, hsa-miR-21-5p, hsa-miR-29b-3p, hsa-miR-126-3p, hsa-miR-192-5p, hsa-miR-223-3p, and hsa-miR-375) widely related to glucose metabolism, IR status, pre-diabetic status and biomarkers of T2D, based on the use of updated reviews and databases (22 ).
3
Circulating RNA Purification from CSF
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Circulating RNA from CSF was purified from 300 or 500 μL of starting material using the mirVana PARIS Isolation kit (Applied Biosystems) according to the manufacturer’s protocol. Briefly, the initial volume of sample (300 μL in most cases) was mixed with the same volume of 2× Denaturing solution containing 375 μL of 2-mercaptoethanol. At this point, two exogenous miRNAs (cel-miR-39 and cel-miR-54) were added at 5 pM to verify the quality of the extraction process. The same volume of acid-phenol:chloroform was then added, and the upper aqueous phase obtained after centrifugation (17,000× g, 10 min, 19 °C) was recovered. This phase was mixed with 100% ethanol and placed into a filter cartridge provided in the kit. After the RNA washing procedures, total RNA was eluted with 40 µL of nuclease-free water and stored at −80 °C for its lateruse.
4
Plasma RNA Extraction and Validation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RNA was extracted from 300 μL of plasma using a mirVana PARIS isolation kit (Applied Biosystems, Vilnius, Lithuania) according to the manufacturer’s instructions. Non-human cel-miR-39 was spiked into the plasma immediately before extraction. RNA isolation efficiency was examined by RT-qPCR quantification of cel-miR-39 to guarantee homogeneous retrotranscription and cDNA synthesis.
5
Extracting total RNA from CSF or plasma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from CSF or plasma using mirVana PARIS Isolation Kit (Applied Biosystems) according to the manufacturer’s instructions. In brief, 300 μl of sample was mixed with an equal volume of 2× Denaturing solution. Spike-in cel-miR-39 and cel-miR-54 exogenous miRNAs were added to check the quality of the extraction. Then, the same volume of acid-phenol:chloroform was added. After centrifugation (17.000×g, 10 min, 19 °C), the upper aqueous phase was recovered and mixed with 100% ethanol. After being placed into a filter cartridge, three washing steps were performed. Total RNA was eluted with 40 μl of nuclease-free water.
6
Extracting Circulating RNAs from Biofluids
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Circulating RNA from cell-free CSF and serum was extracted using a mirVana PARIS Isolation Kit (Applied Biosystems, CA) according to the manufacturer's protocol. Briefly, 300 μL of sample were mixed with an equal amount of ×2 denaturing solution. The same volume of acid-phenol:chloroform was then added, and the upper aqueous phase was recovered after centrifugation at 17,000g for 10 minutes. This recovered phase was mixed with 100% ethanol and placed into a provided filter cartridge. After 3 washing steps, total RNA was eluted with 40 μL of nuclease-free water and stored at −80°C for its posterior use.
7
Plasma RNA Extraction and Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RNA extraction was performed from 300 μL of plasma using a mirVana PARIS isolation kit (Applied Biosystems, Vilnius, Lithuania) according to the manufacturer’s instructions. Non-human cel-miR-39 was spiked into the plasma immediately before extraction. RNA concentration and integrity were examined by RT-qPCR quantification of cel-miR-39. RNU6 was selected as the plasma quality indicator because it is highly expressed in different cell types but not well detected in plasma. High detection of RNU6 in plasma indicates cellular contamination [19 (link)].
8
Robust Extracellular RNA Extraction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Circulating RNA extraction and puri cation Total RNA of CSF was isolated according to the manufacturer's instructions using the mirVana PARIS Isolation Kit (Applied Biosystems). In the procedure, a denaturing solution was diluted 2 to one with 300 µL CSF. An equal amount of acid phenol and chloroform was then mixed. The aqueous phase was collected after centrifugation and combined with 100% ethanol on a lter cartridge. It was then washed thrice with 40 µL of nuclease-free water.
9
Plasma miR-21 in Septic AKI
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Evaluation of miR-21’s role in sepsis-associated kidney dysfunction was carried out by comparing the miR-21 plasma levels between 53 patients with septic AKI and 21 septic patients without AKI. Then, 200 µL of plasma was obtained from each patient for total RNA isolation with the mirVana PARIS isolation kit (Ambion, TX, USA) based on the corporation’s protocols. The 7900HT Fast Real-Time PCR System (Thermo Scientific, MA, USA) was used to quantify miR-21 expression. Informed written consent was obtained from each participant before study commencement. All human investigational procedures were in line with principles, as stated in the Declaration of Helsinki and were approved by the institutional review committee of Harbin Medical University.
10
Serum miRNA Isolation and Quantification
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!