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miRNeasy Serum/Plasma Advanced Kit

For phenol-free isolation of total RNA including miRNA from serum and plasma samples
  • Phenol-free protocol – no need for phase separation or working under the hood
  • MinElute columns allow for small elution volumes (approx. 20 µl)   
  • Consistent, pure RNA for all downstream applications
The miRNeasy Serum/Plasma Advanced Kit enables the isolation of total RNA, including miRNA, from a minimum of 200 µl of sample. The phenol-free protocol allows all steps to be performed on the bench and removes the need for phase separation, making the kit suitable for automation, e.g., on the QIAcube. The kit includes UCP columns that go through a rigorous ultraclean production process to ensure highly pure RNA eluates.
Cat No./ID: 217204
miRNeasy Serum/Plasma Advanced Kit (50)
€426.00
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For 50 total RNA preps: 50 RNeasy MinElute Spin Columns, Collection Tubes (1.5 ml and 2 ml), RNase-Free Reagents and Buffers
The miRNeasy Serum/Plasma Advanced Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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High recovery of miRNA without the need for phenol.
miRNA was isolated from 200 µl plasma from 3 different samples using the miRNeasy Serum/Plasma Advanced Kit (light blue bars) or the miRNeasy Serum/Plasma Kit (dark blue bars). Five microliters of recovered RNA, in a total volume of 25 µl, was used for cDNA synthesis using the miScript system, followed by PCR detection. Both kits show good recovery of miR16.
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Detection of miRNAs preferentially found inside and outside of extracellular vesicles.
miRNA was isolated from 200 µl plasma from 20 different samples using the miRNeasy Serum/Plasma Advanced Kit. Five microliters of recovered RNA, in a total volume of 25 µl, was used for cDNA synthesis using the miScript system, followed by PCR detection of 3 different miRNAs found to be primarily present inside or outside of extracellular vesicle, respectively. The miRNeasy Serum/Plasma Advanced Kit can detect miRNAs both inside and outside of vesicles.

Part of this result have been generated within the CANCER ID project, funded by the Innovative Medicines Initiative 2 Joint Undertaking (JU) under grant agreement No 115749. The JU receives support from the European Union’s Horizon 2020 research and innovation program and EFPIA.
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Competitive yields and performance.
The miRNeasy Serum/Plasma Advanced Kit shows comparable or better performance to kits from other suppliers. miRNA (A) or mRNA (B) was isolated from 200 µl plasma from 2 different samples using the miRNeasy Serum/Plasma Advanced Kit or kits from other suppliers. Depending on the elution volume, an equal amount of the recovered RNA was applied in a total volume of (A) 25 µl for cDNA synthesis using the miScript system, followed by PCR detection of miR122a and miR150 miRNA, respectively, or (B) applied in a total volume of 20 µl for RT-PCR using the QuantiFast SYBR Green Kit and detection of mRNA-RPL12. The miRNeasy Serum/Plasma Advanced Kit shows comparable or better performance compared to kits from other suppliers for both miRNAs.
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miRNeasy Serum/Plasma Advanced Kit procedure.
Performance
The miRNeasy Serum/Plasma Advanced Kit provides a phenol-free alternative to the miRNeasy Serum/Plasma Kit with similar RNA yields and quality (see figure High recovery of miRNA without the need for phenol). The miRNeasy Serum/Plasma Advanced Kit efficiently isolates total RNA, including miRNA, from serum and plasma samples starting from a minimum input amount of 200 µl. The phenol-free protocol uses easy-to-automate MinElute spin column technology without the need for a vacuum pump (see figure miRNeasy Serum/Plasma Advanced Kit procedure). Extracellular vesicles are lysed to allow analysis of RNA in and outside of vesicles (see figure Detection of miRNAs preferentially found inside and outside of extracellular vesicles). Unlike kits from other suppliers, the miRNeasy Serum/Plasma Advanced Kit combines ease-of-use and a phenol-free protocol without any compromise on RNA quality or yield (see figure Competitive yields and performance).
Principle
For analysis of the complete plasma RNA content, including RNA contained within extracellular vesicles, Buffer RPL mixed with the plasma sample is used to ensure efficient lysis. The kit includes Buffer RPP, which precipitates contaminants such as plasma proteins. The kit employs a protocol based on spin-column technology. The sample is first applied to the spin column to bind total RNA and any unwanted analytes or contaminants are removed in subsequent wash and centrifugation steps before the sample is eluted in the final centrifugation step.
Procedure
Serum or plasma (200 µl) are mixed with Buffer RPL to release and stabilize RNA from plasma proteins and extracellular vesicles. The sample is then mixed with Buffer RPP and centrifuged to precipitate proteins. Isopropanol is added to the supernatant to provide the appropriate conditions for RNA molecules (>18 nucleotides) to bind to the silica membrane. The sample is then applied to the RNeasy UCP MinElute spin column, where RNA binds to the membrane and other contaminants are washed away in subsequent wash steps. In the final step, total RNA (>18 nucleotides) is eluted using RNase-free water.
Applications
The miRNeasy Serum/Plasma Advanced Kit isolates total cell-free and exosomal RNA from plasma or serum. Downstream applications include:
  • Biomarker discovery
  • Expression profiling
  • Next-generation sequencing
  • Quantitative, real-time RT-PCR
  • Microarray analysis
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