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miRNeasy 96 Advanced QIAcube HT Kit

For automated purification of total RNA, including miRNA, from serum and plasma samples

  • Phenol-free miRNA isolation from human, mouse or rat serum and plasma samples
  • Simple and reliable automated processing for time and cost savings
  • High-quality miRNA ready for use in any downstream application
  • Consistent miRNA yields from small amounts of starting material

The miRNeasy 96 Advanced QIAcube HT Kit is designed for automated high-throughput purification of cell-free total RNA – primarily miRNA and other small RNA – from small volumes of serum and plasma on the QIAcube HT. Using proven RNeasy silica-membrane technology in a convenient 96-well format, contaminants and inhibitors are removed to yield high-quality RNA ready for downstream analysis.

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Cat No./ID: 217261
miRNeasy 96 Advanced QIAcube HT Kit (5)
$3,395.00
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For 480 preps: RNeasy 96 plates, RNase-free water, buffers
Cat No./ID: 950067
QIAcube HT Plasticware
$331.00
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Plasticware for 480 typical preps on QIAcube HT:
5 S-Blocks, 5 Elution Microtubes RS (EMTR), 120 x 8-Well Strip Caps for EMTR, 9 x 96 Filter-Tips OnCor C, TapePad
The miRNeasy 96 Advanced QIAcube HT Kit is intended for molecular biology applications. This product is neither intended for the diagnosis, prevention, or treatment of a disease, nor has it been validated for such use either alone or in combination with other products.

Product Details

9

Performance comparable with manual kit using equal volumes.

The miRNeasy Advanced QIAcube HT Kit performs comparably with the manual miRNeasy Serum/Plasma Advanced Kit and miRNeasy Kit. miRNA was isolated from 200 µl plasma samples according to each protocol. The elution volume of 100 µl with the QIAcube HT was 5 times greater than the 20 µl eluation volume for the manual kits. Equal volumes of eluate (5 µl) was applied in a total reaction volume of 20 µl for cDNA synthesis, followed by PCR detection of miR150 miRNA, miR16, miR122a, and miR let7a as indicated. The miRNeasy 96 Advanced QIAcube HT Kit performed comparably with the manual kits.

9

Performance comparable with manual kit using equal amounts of RNA.

The miRNeasy Advanced QIAcube HT Kit shows comparable performance to the manual kits miRNeasy Serum/Plasma Advanced kit as well as the miRNeasy Kit. miRNA was isolated from 200 µl plasma samples according to each protocol. Depending on the elution volume (100 µl on the QCHT and 20 µl for the manual kits), an equal amount of the recovered RNA was applied in a total volume of 20 µl for cDNA synthesis, followed by PCR detection of miR150 miRNA, miR16, miR122a, and miR let7a respectively. The miRNeasy Serum/Plasma Advanced Kit shows comparable performance compared to the manual kits. In addition manual handling is less and handling easy.

9
Processing different amounts of plasma.
miRNA was isolated from 50, 100 and 200 µl volumes of plasma from two healthy donors using the miRNeasy Serum/Plasma Advanced Kit and the miRNeasy Advanced QIAcube HT Kit. Detection of miR150 shows good performance even starting with low amounts of plasma (50 µl).
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Compatibility with different blood collection tubes.
Blood from healthy donors was collected using EDTA or serum collection tubes. Plasma or serum was generated and miRNA isolated miRNA from 200 µl using the miRNeasy Advanced QIAcube HT Kit, miRNeasy Serum/Plasma Advanced Kit or miRNeasy Kit. miR150 was detected using 5 µl eluate. The miRNeasy Advanced QIAcube HT Kit performed comparably with the manual kits.
9
Comparable or better yields and performance.
miRNA was isolated from 50–250 µl plasma from 2 different samples according to the manufacturer’s instructions. Equal amounts of recovered RNA were used in a total volume of 25 µl for cDNA synthesis, followed by PCR detection of miR150 miRNA. The miRNeasy 96 Advanced Kit performed comparably or better than kits from other suppliers for both plasma samples.
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miRNeasy 96 Advanced QIAcube HT Kit procedure.

The miRNeasy 96 Advanced QIAcube HT Kit has been designed for phenol-free, rapid medium- to high-throughput purification of cell-free RNA from human and animal serum or plasma on the QIAcube HT instrument. The phenol-free extraction procedure allows cell-free RNA extraction from liquid biopsy samples quickly and with minimal manual interaction.

Performance

The miRNeasy 96 Advanced QIAcube HT Kit is the medium- to high-throughput version of the miRNeasy Serum/Plasma Advanced Kit and provides the same yield and quality of RNA using phenol-free isolation technology (see figures Performance comparable with manual kit using equal volumes) and Performance comparable with manual kit using equal amounts of RNA). The miRNeasy 96 Advanced QIAcube HT Kit efficiently isolates total RNA, including miRNA, from human and animal serum and plasma samples from 50 µl to 200 µl (Processing different amounts of plasma). The kit is compatible with the majority of blood collection devices (see figure Compatibility with different blood collection tubes). The phenol-free protocol uses easy-to-automate spin column technology in 96-well format and minimize hands on time. Unlike kits from other suppliers, the miRNeasy 96 Advanced QIAcube HT Kit combines ease-of-use and a phenol-free protocol without any compromise on RNA quality or yield (see figure Comparable or better yields and performance).

Principle

The miRNeasy 96 Advanced QIAcube HT Kit enables automated purification of cell-free total RNA – primarily miRNA and other small RNA, from 20–200 µl of human or animal plasma using the QIAcube HT instrument. Selective binding properties of a silica membrane are combined with a high-throughput 96-well format for fully automated, simultaneous processing of 24–96 samples on the QIAcube HT instrument. The kit combines guanidine-based lysis of samples, an inhibitor removal step and silica-membrane-based purification of total RNA.

The system provides two protocols, a transfer protocol to move the lysate from the protein precipitation step to the RNeasy 96 plate and the cell-free RNA isolation protocol to extract miRNA from the lysate. The procedures yield high-quality RNA that performs well in downstream applications .

 

Procedure

Lysis is performed manually under highly denaturing conditions with guanidine thiocyanate to immediately inactivate RNases and ensure purification of intact RNA (see figure miRNeasy 96 Advanced QIAcube HT Kit procedure.)

After the run is started, isopropanol is added to the RNA-containing supernatant to provide appropriate binding conditions for all RNA molecules from approximately 18 nucleotides (nt) upwards. The samples are afterwards transferred (manually or automated) to the wells of the RNeasy 96 plate. Total RNA binds and contaminants are efficiently washed away. High-quality RNA is then eluted in RNase-free water, ready for use in any downstream application.

 

Applications

The miRNeasy 96 Advanced QIAcube HT 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

Specifications

Features
Specifications
Elution volume 80–100 µl
Input volume Up to 200 µl
Number of preps per run 24–96 samples (processed in increments of 8)
Run time 96 samples in approximately 93 minutes; 24 samples in approximately 54 minutes

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