Description
Background information
A range of applications from reverse transcription (RT)-PCR to microarray analysis and cDNA library generation require high-quality RNA. Using mRNA instead of total RNA as starting material can significantly improve the sensitivity and specificity of the results. This is particularly important, when the starting material is limited or rare transcripts are analyzed. µMACS and MultiMACS mRNA Isolation Kits enable a highly sensitive mRNA isolation, faster even than traditional total RNA isolations.
Detailed procedure
After cell lysis and clearing of the cell lysate, the mRNA is magnetically labeled with µMACS Oligo(dT) MicroBeads. Due to the small size of the MicroBeads, the hybridization to the poly(A)+ tail of the mRNA molecules is completed within seconds. The sample is loaded onto a MACS Column placed in the magnetic field of a µMACS Separator. After washing, the magnetically labeled mRNA is retained on the column. By applying Elution Buffer on the column, pure mRNA is eluted from the column. The µMACS mRNA Isolation Kits are designed for manual low-throughput mRNA isolation whereas the MultiMACS mRNA Isolation Kits are designed for high-throughput applications in a 96-well format. The MultiMACS Kits are used in combination with the MultiMACS M96 Separator that can be integrated in a robotic system to achieve a fully automated approach.
Overview
The µMACS and MultiMACS mRNA Isolation Kits facilitate fast and efficient mRNA isolation without prior preparation of total RNA. Pure mRNA is directly isolated from cells, tissue, or blood by magnetic labeling with µMACS Oligo(dT) MicroBeads. Due to the fast reaction kinetics the small (50 nm) µMACS Oligo(dT) MicroBeads instantly bind to mRNA molecules and allow for efficient and reproducible isolation of high-purity mRNA. This magnetic bead-based technology ensures reliable results from just a few cells up to 107 cells.
