|
|
 |
|
| Simplify & amplify! |
Gene expression profiling even from a single cell
The µMACS™ SuperAmp™ Kit1 allows extremely sensitive mRNA isolation, cDNA synthesis, and millionfold amplification of the mRNA-derived cDNA by global polymerase chain reaction starting with 1–10,000 cells or appropriate amounts of tissue.
Principle of SuperAmp Technology
Based on the well-established MACS® Technology mRNA is isolated utilizing paramagnetic oligo(dT) MicroBeads. Uniform-sized cDNA fragments are generated by an unique in-column cDNA synthesis procedure. All cDNA fragments are tailed at the 3’-end and are amplified by single-primer global PCR. Finally, the amplified DNA fragments are labeled in a Klenow labeling reaction.
Click here and have a look on a detailed illustration of the Principle of the µMACS™ SuperAmp™ Technology.
The same technology is already successfully applied in Miltenyi Biotec's SuperAmp™ Microarray Service2-8
Any type of rare cells can be used as starting material:
– Cell lines
– Cells sorted with MACS® Technology
– Cells sorted by flow cytometry
– Tissue biopsies
– Laser-capture microdissected cells
The unique SuperAmp™ Technology provides:
• Exceptional sensitivity
Extremely small, superparamagnetic MACS® MicroBeads instantly bind mRNA molecules from small samples, while the MACS® Column Technology simplifies the required washing steps resulting in high-purity mRNA—even from very small sample materials. The unique one-step mRNA isolation and in-column cDNA synthesis procedure reduces loss of material due to tube-to-tube transfer.
• High reproducibility
The carefully optimized reverse transcription (RT) protocol generates small first-strand cDNA fragments of consistent length, reducing PCR bias in the subsequent amplification procedure. In addition, PCR bias is avoided by a single-primer global PCR amplification procedure with uniform annealing conditions for all transcripts.
• Time- and labor-saving
The reliable SuperAmp™ Technology is much faster than 2-rounds T7 amplification: only ten hours hands-on time from lysed cells to amplified, labeled, and purified DNA. |
| SuperAmp Sample Shipment |
The SuperAmp Shipment Buffer Kit was developed for a safe shipment of fragile samples for subsequent amplification of mRNA with the µMACS SuperAmp Kit.
The SuperAmp Shipment Buffer provided with the SuperAmp Shipment Buffer Kit allows a very efficient cell lysis and stabilizes the fragile samples for shipment. |
| Columns |
| µ Columns (included in the kit) |
| |
| Figure 1 |
| Reliability of the µMACS™ SuperAmp™ Protocol using cell lines. A serial dilution of 1–10,000 Jurkat cells was generated and each dilution was independently processed using the µMACS™ SuperAmp™ Kit. The amplified and Cy5-labeled cDNA from Jurkat cells was hybridized in a two-color microarray experiment against amplified and Cy3-labeled cDNA from 1000 Raji cells. Each experiment was repeated thrice and the independently amplified samples were hybridized on PIQOR™ Immunology Microarrays. A: Pearson correlation coefficients (r) of independent repeats. B: Hierarchical clustering with no obvious grouping of repeats or cell numbers. |
 |
|
| Figure 2 |
| Reliability of the µMACS™ SuperAmp™ Protocol using primary cells. CD14+ cells were separated from 5 x 107 peripheral blood mononuclear cells to a purity of 98% using MACS® Technology. A serial dilution of 5-10,000 CD14+ cells was generated and each dilution was independently processed using the µMACS™ SuperAmp™ Kit. The amplified and Cy5-labeled cDNA from CD14+ cells was hybridized in a two-color microarray experiment against amplified and Cy3-labeled cDNA from 1000 Raji cells. Each experiment was repeated thrice and the independently amplified samples were hybridized on PIQOR™ Immunology Microarrays. A: Pearson correlation coefficients (r) of the three independent repeats. B: Hierachical clustering with no obvious grouping of repeats or cell numbers. |
 |
|
|
|
|
| My favorites |
| Your current favorite list is empty. |
|
| Products |
| μMACS SuperAmp Kit |
- for 10 reactions
Components
- 1.25 mL Lysis/Binding Buffer
- 20 µL tRNA Solution
- 50 µL µMACS™ SuperAmp™ MicroBeads
- 4 mL Wash Buffer
- 4 mL Equilibration/Wash Buffer
- 0.2 mL Resuspension Buffer C
- 10 µL µMACS™ Sealing Solution
- 2×1 mL Tailing Wash Buffer
- 1 mL Resuspension Buffer T
- 0.6 mL Resuspension Buffer P
- 0.22 mL Double Distilled Water
- 0.3 mL Resuspension Buffer K
- 10 × Lyophilized First-strand cDNA Mix
- 20 × Lyophilized Tailing Mix
- 10 × Lyophilized PCR Mix
- 10 × Lyophilized Klenow Mix I
- 10 × Lyophilized Klenow Mix II
- 10 × µ Columns
- 20 × 200µL Tubes
Download datasheet
130-093-242
|
| μMACS SuperAmp Starting Kit |
- for 10 reactions
Components
- 1 × µMACS™ SuperAmp™ Kit
- 1 × thermoMACS™ Separator
- 1 × MACS® Multistand
130-093-251
|
| SuperAmp Shipment Buffer Kit |
- For the shipment of 10 SuperAmp samples
Components
- 70 μL SuperAmp Shipment Buffer
- 10× 0.2mL SuperAmp Shipment Tubes
- 1× SuperAmp Shipment Tube Container
Download datasheet
130-095-179
|
|
| References |
| 1. Biagioli et al. (2009) PNAS 106(36): 15454-15459 |
| 2. Stehling-Sun et al. (2009) Nature Immunol. 10(3): 289-296. |
| 3. Paulsen et al. (2009) J. Neurosci. Methods 177(1): 87-93. |
| 4. Li et al. (2009) Differentiation 77(1): 95-102 |
| 5. Hardt et al. (2008) Mol. Cell. Neurosc. 39: 418-428. |
| 6. Auffray et al. (2007) Science 317: 666-670. |
| 7. Appay et al. (2007) Journal of Immunology 179: 7406-7414. |
| 8. Zeytun et al. (2007) Adv. Exp. Med. Biol. 598: 342-357. |
|
|
|
© 1999 - 2010 Miltenyi Biotec ¦ Terms & conditions ¦ Legal notes
|
