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RNeasy 96 QIAcube HT Kit

For automated high-throughput isolation of RNA from animal and human cells and tissue

  • 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

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.

Артикул: 950067
QIAcube HT Plasticware
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
Артикул: 74171
RNeasy 96 QIAcube HT Kit (5)
For 480 preps: RNeasy 96 plates, RNase-free water, buffers

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.

This economical, easy-to-use system provides the same results as other trusted QIAGEN purification technologies. 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.

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).

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 
* Amounts can vary due to factors such as species, developmental stage, and growth conditions. If the RNeasy procedure enriches for RNA >200 nucleotides, the total RNA yield does not include 5S rRNA, tRNA, and other low molecular weight RNAs, which make up 15–20% of total cellular RNA.

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.

Specifications for purification of total RNA from cells

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 ?l Buffer RLT
Elution volume 110 ?l
Duration 96 samples in approximately 93 or 114* minutes
24 samples in approximately 54 or 73* minutes
* Including optional DNase digestion.

Specifications for purification of total RNA from tissues

Specification Description
Number of samples 24–96 samples (to be processed in increments of 8)
Sample input volume 350 ?l aqueous phase from up to 40 mg frozen or 20 mg
stabilized tissue homogenized in 750 ?l QIAzol Lysis Reagent
Elution volume 110 ?l
Duration 96 samples in approximately 73 or 93* minutes
24 samples in approximately 34 or 54* minutes
* Including optional DNase digestion.

Specifications for purification of total RNA, including miRNA, from cells or tissue

Specification Description
Number of samples 24–96 samples (to be processed in increments of 8)
Sample input volume 350 ?l aqueous phase from up to 40 mg frozen or 20 mg
stabilized tissue homogenized in 750 ?l QIAzol Lysis Reagent
Elution volume 110 ?l
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.

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