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RNeasy 96 QIAcube HT Kit (5)
Cat. No. / ID: 74171
For 480 preps: RNeasy 96 plates, RNase-free water, buffers
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RNeasy 96 QIAcube HT Kit
QIAcube HT Plasticware
The RNeasy 96 QIAcube HT Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
✓ 24/7 automatic processing of online orders
✓ Knowledgeable and professional Product & Technical Support
✓ Fast and reliable (re)-ordering
Features
- Simple and reliable automated processing for time and cost savings
- Flexible procedures for isolating total RNA or total RNA, including miRNA
- Protocols for various starting materials and desired size distribution
- High-quality purified RNA ready for use in any downstream application
- Consistent RNA yields from small amounts of starting material
Product Details
The RNeasy 96 QIAcube HT Kit enables simple, automated purification of total RNA or total RNA, including miRNA, from animal and human cells and tissue samples on the QIAcube HT system. 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.
Performance
Purification of RNA on QIAcube HT is simple and reliable with the RNeasy 96 QIAcube HT Kit and provides linear sample recovery with no significant well-to-well variation across a 96-well plate (see figure Linear RNA recovery and Reproducible 96-well RNA purification).
This economical, easy-to-use system provides the same results as other trusted QIAGEN purification technologies (see figure Automated purification of RNA from tissues). Analysis of QIAcube HT purified samples shows high RNA Integrity Scores (RIS) when analyzed with the QIAxcel Advanced system, and performance similar to other automated QIAGEN purification solutions (see figure High-quality RNA from tissues).
The RNeasy 96 QIAcube HT Kit provides consistent yields from small amounts of starting sample material (see table Typical total RNA yields from cells with the RNeasy 96 QIAcube HT Kit).
This economical, easy-to-use system provides the same results as other trusted QIAGEN purification technologies (see figure Automated purification of RNA from tissues). Analysis of QIAcube HT purified samples shows high RNA Integrity Scores (RIS) when analyzed with the QIAxcel Advanced system, and performance similar to other automated QIAGEN purification solutions (see figure High-quality RNA from tissues).
The RNeasy 96 QIAcube HT Kit provides consistent yields from small amounts of starting sample material (see table Typical total RNA yields from cells with the RNeasy 96 QIAcube HT Kit).
| Cell line | Source | RNA yield (µg per 105 cells)* |
|---|---|---|
| HeLa | Human cervical carcinoma | 1.6 |
| LMH | Chicken hepatoma | 1.3 |
| COS-7 | Monkey kidney, SV-40 transformed | 3.1 |
| Huh7 | Human hepatoma | 2.0 |
| Jurkat | Human T-cell leukemia | 1.4 |
| K-562 | Human chronic myelogenous leukemia in blast crisis | 1.9 |
See figures
Principle
The RNeasy 96 QIAcube HT Kit enables automated purification of total RNA or total RNA, including miRNA, from 5 x 105 animal or human cells or <40 mg tissue using the QIAcube HT instrument. Six instrument protocols are available, depending on the starting material and desired size distribution of purified RNA. Cell samples can be purified directly using the RNeasy 96 QIAcube HT Kit and the dedicated kit protocol. Tissue samples (<40 mg) and samples for purification of small RNA are processed following sample lysis in QIAzol Lysis Reagent. The procedures yield high-quality RNA that performs well in downstream applications.
The RNeasy 96 QIAcube HT Kit combines the selective binding properties of a silica-based membrane with a high-throughput 96-well format, and is designed for fully-automated, simultaneous processing of 24–96 samples on the QIAcube HT instrument.
| Specification | Description |
|---|---|
| Number of samples | 24–96 samples (to be processed in increments of 8) |
| Sample input volume | Up to 5 x 105 cells, homogenized in 140 μl Buffer RLT |
| Elution volume | 110 μl |
| Duration | 96 samples in approximately 93 or 114* minutes 24 samples in approximately 54 or 73* minutes |
| Specification | Description |
|---|---|
| Number of samples | 24–96 samples (to be processed in increments of 8) |
| Sample input volume | 350 μl aqueous phase from up to 40 mg frozen or 20 mg stabilized tissue homogenized in 750 μl QIAzol Lysis Reagent |
| Elution volume | 110 μl |
| Duration | 96 samples in approximately 73 or 93* minutes 24 samples in approximately 34 or 54* minutes |
| Specification | Description |
|---|---|
| Number of samples | 24–96 samples (to be processed in increments of 8) |
| Sample input volume | 350 μl aqueous phase from up to 40 mg frozen or 20 mg stabilized tissue homogenized in 750 μl QIAzol Lysis Reagent |
| Elution volume | 110 μl |
| Duration | 96 samples in approximately 78 minutes 24 samples in approximately 38 minutes |
Procedure
Cell lysis is performed manually under highly denaturing conditions with guanidine thiocyanate to immediately inactivate RNases and ensure purification of intact RNA. After the run is started, ethanol is added to the lysate to provide appropriate binding conditions, and the samples are then applied to the wells of the RNeasy 96 plate. Total RNA binds and contaminants are efficiently washed away. High-quality RNA is then eluted in a small volume of RNase-free water, ready for use in any downstream application.
With the automated procedure for purification of total RNA from animal or human cells, all RNA molecules longer than 200 nucleotides are purified. This provides enrichment for mRNA, since most RNAs <200 nucleotides (such as 5.8S rRNA, 5S rRNA, and tRNA, which together comprise 15–20% of total RNA) are selectively excluded. The size distribution of the purified RNA is comparable to that obtained by centrifugation through a CsCl cushion, where small RNA does not sediment efficiently.
For purification of total RNA from tissue, or of total RNA including small RNA, from cells and tissues, we recommend first using QIAzol Lysis Reagent as the lysis buffer. Detailed procedures are provided in the kit handbook.
With the automated procedure for purification of total RNA from animal or human cells, all RNA molecules longer than 200 nucleotides are purified. This provides enrichment for mRNA, since most RNAs <200 nucleotides (such as 5.8S rRNA, 5S rRNA, and tRNA, which together comprise 15–20% of total RNA) are selectively excluded. The size distribution of the purified RNA is comparable to that obtained by centrifugation through a CsCl cushion, where small RNA does not sediment efficiently.
For purification of total RNA from tissue, or of total RNA including small RNA, from cells and tissues, we recommend first using QIAzol Lysis Reagent as the lysis buffer. Detailed procedures are provided in the kit handbook.
Applications
The RNeasy 96 QIAcube HT Kit provides efficient, high-throughput RNA sample preparation for research use in fields such as drug screening and basic research. The purified RNA is ready to use in any downstream application, including:
- RT-PCR and qRT-PCR
- Differential display
- cDNA synthesis
- Northern, dot, and slot blot analyses
- Primer extension
- Poly A+ RNA selection
- RNase/S1 nuclease protection
Supporting data and figures
Automated purification of RNA from tissues.
RNA was isolated from 10 mg of various rat tissues stabilized in RNAlater RNA Stabilization Reagent. RNA was purified using QIAzol Lysis Reagent and the QIAcube HT, BioRobot Universal System, or QIAcube. Purified RNA was analyzed by qRT-PCR for pgk1 mRNA using the QuantiFast Probe RT-PCR Kit. CT values indicate similar performance of RNA extracted with the different purification platforms.
Resources
Protocol Files (12)
Safety Data Sheets (1)
Technical Information (2)
Quick-Start Protocols (4)
Kit Handbooks (2)
Instrument User Manuals (1)
Certificates of Analysis (1)
FAQ
Can I run the QIAcube HT with the QIAxtractor software or vice versa?
For what batch size is the QIAcube HT Plasticware designed?
Can I upgrade a Corbett CAS-1820 to a QIAcube HT?
What file is required for reporting a QIAcube HT or QIAxtractor issue?
I am getting vacuum errors on my QIAcube HT. What should I do?
1. Check the vacuum tubing to make sure it is not pinched or obstructed in some way. Sometimes the tubing can become pinched against the bench top when the vacuum station is placed too far from the instrument.
2. Make sure that the channeling adapter is properly placed. Check if the carriage is moving the plate completely from the waste chamber position to the elution chamber position. If the calibration seems incorrect, the instrument may need to be re-taught at this position.
3. When using less than a full plate, make sure all the unused columns are covered with an adhesive seal.
Also, make sure each column of 8 samples is full. When processing a number of samples that results in < 8 samples per column, use water as a substitute in the empty positions until the column is filled. This ensures even vacuum across all unsealed well positions.
4. Make sure correct plates are used for the selected protocol.
5. When the vacuum is on, please listen for any hissing noise from the tubing that is coming down to the waste container. This could indicate leaks in the tubing.
If the issue persists, please contact QIAGEN Technical Service.
FAQ ID -9106
What do I do if the QIAcube HT or QIAxtractor software runs in virtual mode while the instrument is connected to the computer?