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.