The protocol for collection of peripheral blood samples was approved by the Institutional Review Board at the University of Alabama at Birmingham, and all donors provided written, informed consent. Blood was collected into EDTA-tubes. Within 30 min of collection, the plasma was isolated (∼5 ml) and stored at −80°C. One milliliter plasma was centrifuged at 14 000 relative centrifugal force for 15 min and total RNA was isolated from the supernatant using the Plasma/Serum Circulating and Exosomal RNA Purification Kit (Slurry Format) (Norgen Biotek) following the manufacturer's directions. The eluate from this kit was further concentrated using the RNA Clean-Up and Concentration Kit (Norgen Biotek) using 20 μl elution buffer to collect the RNA.
Plasma serum circulating and exosomal rna purification kit slurry format
Manufactured by Norgen Biotek
Sourced in Canada
The Plasma/Serum Circulating and Exosomal RNA Purification Kit (Slurry Format) is a lab equipment product designed for the isolation and purification of circulating and exosomal RNA from plasma or serum samples. It utilizes a slurry-based approach to ensure efficient extraction of RNA.
Automatically generated - may contain errors
Lab products found in correlation
Rna clean up and concentration kit, by Norgen Biotek (2 mentions)
Rnalater, by Thermo Fisher Scientific (2 mentions)
Mirna purification kit, by Norgen Biotek (1 mentions)
Miseq reagent nano kit, by Illumina (1 mentions)
Pippin prep with a 3 agarose gel cassette, by Sage Science (1 mentions)
Agilent high sensitivity dna kit, by Agilent Technologies (1 mentions)
Hiseq 4000, by Illumina (1 mentions)
Dna clean concentrator 5 kit, by Zymo Research (1 mentions)
Agilent rna 6000 pico kit, by Agilent Technologies (1 mentions)
Total exosome isolation reagent, by Thermo Fisher Scientific (1 mentions)
Quant it ribogreen rna assay, by Thermo Fisher Scientific (1 mentions)
7 protocols using plasma serum circulating and exosomal rna purification kit slurry format
1
Plasma RNA Isolation Protocol
2
Plasma RNA Isolation for Sequencing
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Peripheral blood sample collections for the isolation of RNA from human plasma were performed as previously described (22 (link)) in accordance with the Institutional Review Board at the University of Alabama at Birmingham. Briefly, each collection 5 ml of blood was drawn, centrifuged at 2200 g for 10 min to isolate plasma and then stored at −80°C until further use. Total RNA was isolated from 1 ml thawed plasma using the Plasma/Serum Circulating and Exosomal RNA Purification Kit (Slurry Format) (Norgen Biotek) and concentrated to 20 μl using the RNA Clean-Up and Concentration Kit (Norgen Biotek). To limit sample variation between MAD-DASH replicate groups, plasma RNA from multiple donors was combined prior to library construction.
3
Plasma RNA Extraction and Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from (300 µL) plasma using Plasma/Serum Circulating and Exosomal RNA Purification Kit (Slurry Format) (Norgen®, Thorold, ON, Canada, Cat. 42800). Samples were spiked with cel-miR-39-3p 2.7−4 µM (IDT). RNA was eluted in 80 µL of elution buffer.
4
Extraction and Purification of RNA from Solid Tumors and Serum
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
From MSKCC's prospectively maintained tissue bank, fresh frozen tumors or formalin fixed, paraffin embedded (FFPE) tissues mounted on glass slides were obtained for each patient. Solid tumor samples were flash frozen, preserved in RNAlater (Thermofisher AM7024) to minimize RNA degradation, cut and measured to roughly the same size, and grounded with disposable pestles (Sigma Z359947) before being subjected to RNA extraction using the Norgen miRNA purification kit (21300) per manufacturer's instructions. Extracted RNA was then bioanalyzed for integrity prior to library preparation. To extract RNA from FFPE tissue slides, a pathologist (J.S.) reviewed the H&E slides for visual confirmation of tumor cells, and areas containing tumor cells were encircled for extraction. Tumor cells were scraped from corresponding unstained slides with a surgical blade, and miRNAs were isolated as described above. For the serum cohort, RNA was extracted from 1mL of serum using Norgen Plasma/Serum Circulating and Exosomal RNA Purification Kit (Slurry Format) (42800).
5
Serum exRNA Isolation and Small RNA-seq
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
exRNA was isolated from each serum sample using the Plasma/Serum Circulating and Exosomal RNA Purification Kit (Slurry Format) (Norgen Biotek Corp., Ontario, Canada). Quality control of isolated RNA was performed using the Agilent RNA 6000 Pico Kit (Agilent, Santa Clara, CA). Small RNA-seq libraries were constructed using the NEB-Next Multiplex Small RNA Library Prep Set for Illumina (New England Biolabs Inc., Ipswich, MA). The libraries were then cleaned using the DNA Clean & Concentrator-5 Kit (Zymo Research, Irvine, CA), and quality control of the libraries was performed using the Agilent High Sensitivity DNA Kit (Agilent, Santa Clara, CA). Equal volumes of the libraries were pooled, size-selected for products that were 120 to 135 base pairs in length using a Pippin Prep with a 3% agarose gel cassette (Sage Science, Beverly, MA), and run on a MiSeq instrument at the UC San Diego Institute for Genomic Medicine (IGM) Genomics Core using the MiSeq Nano Reagent Kit (Illumina, San Diego, CA). Samples that produced adequate numbers of miRNA read counts were then rebalanced to produce similar numbers of miRNA reads and sequenced on a HiSeq 4000 instrument to produce 1 × 75 bp reads (Illumina, San Diego, CA) at the UC San Diego IGM Genomics Core.
6
Exosomal RNA Extraction from Cell Media
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The exosomes released from cells cultured in SC and DDW cell culture medium without 5-FU or oxaliplatin were precipitated from 4 ml cell culture media using Total Exosome Isolation Reagent (Thermo Fisher Scientific, Inc.) according to the manufacturer's protocol. Subsequently the precipitated exosomal fraction was used for extraction of total RNA using a Plasma/Serum Circulating and Exosomal RNA Purification kit (Slurry Format; Norgen Biotek Corp., Thorold, ON, Canada) for the cells maintained in SC and those maintained in DDW-prepared medium.
7
Extraction and Quantification of Circulating RNA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from Fraction 7 using a Plasma/Serum Circulating and Exosomal RNA Purification Kit (Slurry Format; Norgen Biotek Corp., Thorold, ON, Canada) according to the manufacturer’s instructions. The RNA concentration was measured with Quant-iT Ribogreen RNA Assay (Thermo Fisher, Waltham, Mass).
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!