TurboCapture mRNA Kits

For rapid, easy, high-throughput mRNA purification from cells

S_1301_GEF_RNA0147

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TurboCapture 96 mRNA Kit (5)

Cat. No. / ID:  72251

5 x TurboCapture 96 mRNA Plates, and RNase-Free Buffers
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KitBuffer
TurboCapture mRNA Kit
Buffer TCL
Plate type
96 well
384 well

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • Plate-based mRNA purification in 96-well or 384-well format
  • Few protocol steps and minimal hands-on time
  • mRNA purification and cDNA synthesis in the same well
  • Standard plates compatible with robotic workstations

Product Details

The TurboCapture System uses unique oligonucleotide immobilization technology to provide a fast and simple procedure for high-throughput mRNA extraction. Cell lysates are added to the wells of a TurboCapture plate (96 or 384 wells), and mRNA is allowed to hybridize to the immobilized oligo-dT in each well. Contaminants are washed away, and the isolated mRNA is then either used directly in cDNA synthesis or eluted for use in downstream applications.

Performance

TurboCapture mRNA Kits provide rapid and easy purification of mRNA from cultured cells, including adherent and suspension cells (see figure " mRNA purification from suspension cells"). Due to the standard plate formats and simple workflows, TurboCapture mRNA Kits can be automated, allowing mRNA purification, cDNA synthesis, and PCR to be performed in the same well. This avoids the need for sample transfer and ensures high well-to-well reproducibility (see figure " High well-to-well reproducibility"). When immobilized oligo-dT in the well is used as primer, the synthesized cDNA is covalently linked to the well and can be reused several times (see figure " Sequential PCR analysis"). The kits are ideally suited for high-throughput gene expression analysis applications, such as screening of siRNAs by real-time RT-PCR (see figure " Parallel analysis of siRNAs"). TurboCapture mRNA Kits also provide high sensitivity in gene expression analysis, allowing detection of targets from as little as a single cell (see figure " Gene expression analysis").

See figures

Principle

Rapid, cost-effective plate-based mRNA extraction from adherent and suspension cells, as well as from total RNA, is achieved with minimal hands-on time using the unique oligonucleotide immobilization technology in TurboCapture mRNA Kits. The kits allow mRNA to be purified by hybridizing to immobilized oligo-dT in each well of a 96- or 384-well plate. The immobilized oligo-dT in the well can be used as primer so that cDNA is synthesized while the mRNA remains hybridized, yielding cDNA that is covalently linked to the well. TurboCapture plates are compatible with most thermal cyclers, allowing mRNA hybridization, and subsequent cDNA synthesis and PCR, to take place in the same plate, minimizing sample handling and reducing consumable costs. The procedure can be automated on robotic instruments, due to the standard plate formats and simple workflows.

Procedure

With TurboCapture mRNA Kits, mRNA can be purified from total RNA, as well as from a range of cell types, including adherent and suspension cells. (If purifying mRNA from suspension cells, Buffer TCL, 2x [cat. no. 1031586] must also be purchased.) Simply add sample lysates to the wells of a TurboCapture plate (96 or 384 wells). Allow mRNA to hybridize to the immobilized oligo-dT in each well, and wash away the contaminants. The isolated mRNA can be used directly in cDNA synthesis or, alternatively, can be eluted for use in downstream applications (see flowchart " TurboCapture mRNA procedure"). The immobilized oligo-dT in the well can be used as a primer for cDNA that will be covalently linked to the well and can be reused. Alternatively, soluble oligo-dT primers or random hexamers can be used to prepare soluble cDNA (see figure " mRNA purification and cDNA synthesis in the same well").

See figures

Applications

TurboCapture mRNA Kits are ideally suited for high-throughput gene expression analysis applications such as:

  • Real-time RT-PCR
  • Quantitative real-time RT-PCR
  • Screening of siRNAs by real-time RT-PCR

TurboCapture mRNA Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

 

Comparison of TurboCapture mRNA Kits
Features TurboCapture 96 mRNA Kit TurboCapture 384 mRNA Kit
Applications PCR, cDNA synthesis PCR, cDNA synthesis
Elution volume 80 µl 20 µl
Format 96-well plate 384-well plate
Main sample type Cultured cells Cultured cells
Processing Manual (centrifugation) Manual (centrifugation)

Purification of total RNA, miRNA,

poly A+ mRNA, DNA or protein

mRNA mRNA
Sample amount 80 µl 20 µl
Stabilization Yes Yes
Technology Oligonucleotide immobilization technology Oligonucleotide immobilization technology
Time per run or per prep 30 minutes – 1.5 hours 30 minutes – 1.5 hours
Yield Varies Varies

Supporting data and figures

Resources

Kit Handbooks (1)
For rapid and easy mRNA purification from cultured cells in high-throughput applications
Brochures & Guides (1)
Certificates of Analysis (1)

FAQ

Is mRNA isolation necessary for sensitive RT-PCR?
Usually not. Since RT-PCR is extremely sensitive, as little as 10–200 ng total RNA is sufficient for each 25–50 µl reaction mix, depending on the RT system. For abundant mRNA species, it is possible to use even less than 10 ng total RNA. For rare mRNA species, use a sequence-specific primer in the RT reaction to increase sensitivity. RNA content in various cells and tissues can be found here.
FAQ ID -111
Is the TurboCapture Microplate Holder Type B compatible with the Biomek FX robotic workstation?

This depends on what type of Automated Labware Positioners (ALP) the Biomek FX has. If it is a standard ALP (meaning, a platform with no plate holders on top), then MicroPlate Holder Type A should be used. If there is a detachable plate holder on top of the ALP, then the Microplate Holder Type B should be used.

MicroPlate Holders Typa A and Type B are used to support TurboCapture plates of the TurboCapture 96 mRNA and TurboCapture 384 mRNA Kits for rapid and easy mRNA purification from cultured cells.

FAQ ID -1040
How can I tell TurboCapture Microplate Holder Type A from Type B?

The TurboCapture Microplate Holder Type A is non-detachable, while the Microplate Holder Type B has a detachable base.

Both holders are used to support TurboCapture plates of the TurboCapture 96 mRNA and TurboCapture 384 mRNA Kits for liquid handling on robotic workstations. 

FAQ ID -1039
What is the difference between TurboCapture Microplate Holders Type A and Type B?

TurboCapture MicroPlate Holder Type A has dimensions based on SBS specifications (Society for Biomolecular Screening). Its format is universal and is compatible with a wide range of robotic workstations.

TurboCapture MicroPlate Holder Type B is designed for use with the Biomek 2000. The Microplate Holder Type A has a non-detachable plate-holder, and the Microplate Type B has a detachable plate holder.

 

Both holders are used to support TurboCapture plates of the TurboCapture 96 mRNA and TurboCapture 384 mRNA Kits for rapid and easy mRNA purification from cultured cells.

FAQ ID -1038