MultiMACS™ mRNA Isolation Kits ( #130-092-520 #130-092-519 )

MultiMACS™ mRNA Isolation Kits

High-throughput direct mRNA isolation – from cells or tissue with superparamagnetic μMACS™ MicroBeads.

Flexible format – eight to 96 parallel purifications are performed either semi-automated on the bench or integrated into a liquid-handling system.

Sensitive technique for improved downstream results – with messenger RNA of high yield and purity.

Reliable and convenient processes – Ready-to-use reagents and MACS® Column Technology reduce pipetting steps, sample losses, and hands-on time.

The MultiMACS™ mRNA Isolation Kits provide a robust and reproducible high-throughput mRNA isolation procedure based on MACS® Technology. The core components are the superparamagnetic Oligo(dT) MicroBeads, the MultiMACS Separation Unit, and the Multi-8/96 Columns. The Multi-8/96 Columns are placed into a MultiMACS Separator to provide a strong magnetic field and enable gravity-flow, magnetic mRNA isolation. When a cell lysate, after addition of the Oligo(dT) MicroBeads, is applied to the Multi-8/96 Columns, the MicroBead-labeled mRNA is retained in the magnetic field. Contaminants like ribosomal RNA (rRNA) and DNA are washed away, and hiqh-quality mRNA can be eluted.

96 isolations in 45 minutes . With the MultiMACS mRNA Isolation Kit, 96 mRNA preparations can be performed in 45 minutes directly from cells or tissues. The extremely small MicroBeads, approx. 50 nm in diameter, do not sediment and allow very fast reaction kinetics without further incubation for mRNA hybridization.

Up to 10⁷ and down to a few cells only. The efficient isolation with MACS MicroBeads and the effective washing steps minimize DNA or rRNA contamination to yield highly pure RNA. This not only applies for small cell samples but also for tissues and larger samples up to 10⁷ cells or 30 mg animal tissue. When prior clearing of the cell or tissue lysate is required, gravity-flow Multi-8 and Multi-96 Filters are used in the same setting. The flexible MultiColumn and MultiFilter format allows to process from eight to up to 96 samples in one experiment.

Sophisticated technology. By using gravity flow columns and filters, MultiMACS mRNA Isolation is performed without vacuum manifolds or centrifuges. This also allows full robotic handling when the MultiMACS Separator is placed in a liquid handling system. Therefore, Multi-8/96 Columns and Filters comply with the standard microtiter plate-format. MACS Column Technology avoids time-consuming removal of supernatant as well as bead-carryover.

High-quality messenger RNA. Using MultiMACS mRNA Isolation Kits, variations of yield and real-time PCR cycle thresholds are generally lower (fig. 1 and 2). The gravity-flow column technology additionally prevents cross contamination (fig. 3). Overmore, compared to standard 96-well total RNA isolation kits, genomic DNA contamination is significantly reduced.

Columns

Multi-8 or Multi-96 Columns (included in kit)

Further information

RNA research brochure
[PDF; 2,4 MB]
Isolation of mRNA from stablized whole blood
[PDF; 111,3 KB]
Average RNA yields
[PDF; 66 KB]

Figure 1

Quantification of 96 mRNA samples purified from 100 μg total RNA, mouse liver, with the MultiMACS mRNA Isolation Kit using the MultiMACS Separator. Average yield: 1.3 μg mRNA; coefficient of variation of 96 samples: 5.2%.

Figure 2

Quantitative RT-PCR of 96 samples purified from 100 μg total RNA, mouse liver, with the MultiMACS mRNA Isolation Kit using the MultiMACS Separator. After reverse transcription, quantitative amplification of the housekeeping gene GAPDH was performed (A). Mean cycle threshold: 12.0; coefficient of variation: 2.0%. Display of the dissociation curves of all 96 samples shows pure peaks (B).

Figure 3

No cross contamination visible. RT-PCR products were generated from 1 μL eluate using GAPDH primers (35 cycles) and then separated by gel electrophoresis in a 2% agarose gel. mRNA was purified with the MultiMACS mRNA Isolation Kit using the MultiMACS Separator in a chessboard pattern: 15 mg mouse liver alternated with negative controls containing Lysis/Binding Buffer. The first row shows MultiMACS Separator positions A1–12, B12–1 of the MultiMACS-96 Columns; the second row C1–12, D12–1; the third row E1–12, F12–1; and the fourth row G1–12, H12–1, respectively. Molecular weight markers are in the first and last lane of every row.

<b>No cross contamination visible. </b>RT-PCR products were generated from 1 μL eluate using GAPDH primers (35 cycles) and then separated by gel electrophoresis in a 2% agarose gel. mRNA was purified with the MultiMACS mRNA Isolation Kit using the MultiMACS Separator in a chessboard pattern: 15 mg mouse liver alternated with negative controls containing Lysis/Binding Buffer. The first row shows MultiMACS Separator positions A1–12, B12–1 of the MultiMACS-96 Columns; the second row C1–12, D12–1; the third row E1–12, F12–1; and the fourth row G1–12, H12–1, respectively. Molecular weight markers are in the first and last lane of every row.