mRNA in 15 minutes - Pure mRNA is isolated directly from cells, tissue, or blood, by effective magnetic labeling with extremely small μMACS™ Oligo(dT) MicroBeads. Prior preparation of total RNA is not necessary.
Premium quality - Magnetically labeled mRNA is retained in the MACS® Column while contaminating proteins, genomic DNA, and ribosomal RNA are effectively washed away. This results in reproducible high yields of pure mRNA.
Convenient and safe - The unique MACS Column Technology and ready-to-use, non-toxic reagents facilitate the isolation of intact mRNA even from small samples or challenging material. |
Sensitive gene expression profiles. A range of applications from reverse transcription (RT)-PCR to microarray analysis and cDNA library generation require high-quality RNA. Although the focus of interest is messenger RNA, which is with 1–5% a minor part of total RNA, often total RNA is used, as traditional mRNA isolation methods are time-consuming and cumbersome.
However, starting with messenger RNA can significantly improve the sensitivity and specificity of the results. This is particularly important, when starting material is limited or rare transcripts are analyzed. µMACS mRNA Isolation Kits enable a highly sensitive mRNA isolation faster than traditional total RNA isolations.
Direct isolation. MACS Technology allows high-quality mRNA isolation directly from various biological materials. This includes fresh1,2, frozen, MACS Column-isolated1,2, or cultured cells3, animal or plant tissues3-8, as well as whole blood or total RNA1,9. If a two-step mRNA isolation from biological samples is favored, µMACS mRNA isolation is the method of choice to obtain high-quality mRNA from total RNA.
Unique MACS Column Technology. After cell lysis and clearing of the cell lysate, the mRNA is magnetically labeled. Due to the small size of the µMACS Oligo(dT) MicroBeads (diameter 50 nm), the hybridization to the poly(A)+ tail of the mRNA molecules is completed within seconds.
The superparamagnetic MACS MicroBeads efficiently label the mRNA molecules in a cell lysate, including mRNAs with shortened poly(A) tails. Once magnetically retained in the MACS Column placed in the separator, the mRNA is effectively washed by simply adding the provided buffers.
Thus, virtually no proteins, ribosomal RNA, or genomic DNA are present in the mRNA isolated by MACS Technology. Avoiding multiple pipetting steps, MACS Column Technology minimizes loss of material. Finally, pure mRNA is eluted in a small volume convenient for further processing, while the MicroBeads remain in the column.
The µMACS mRNA isolation runs without toxic chemicals - neither guanidinium-isothiocyanate lysis nor phenol extraction is necessary. Downstream enzymatic treatments as well as removal of genomic DNA traces can be performed directly in the µ Column. For in-column first-strand cDNA synthesis and labeling, please refer to µMACS One-step cDNA Kit or µMACS One-step cDNA Labeling Kit, respectively. |
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| Figure 2 |
| Elution diagramm of mRNA isolated with μMACS Technology (red) or latex resin (green) from 200 μg total RNA of mouse spleen (Bioanalyzer 2100, Agilent Technologies). |
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| Figure 3 |
| mRNA isolated from cells or tissue. mRNA was directly isolated from human peripheral blood lymphocytes (lane 1), monocytes isolated with CD14 MicroBeads (2), hybridoma cells (3), CHO cells (4), mouse spleen (5), mouse lung (6), mouse kidney tissue (7), and from leaves of Forsythia viridissima (8). M: RNA marker. |
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| Figure 4 |
| RT-PCR of mRNA isolated from whole blood. An 838 bp fragment of ß-actin was amplified in an one-step RT-PCR using mRNA isolated from 200 μL (lane 1), 100 μL (3), or 50 μL (5) whole blood. As a negative control, PCR was performed with the respective mRNA preparations in the absence of reverse transcriptase (2, 4, and 6). |
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