Abstract for PolyATtract® mRNA Isolation Systems
The PolyATtract® mRNA Isolation Systems utilize Promega's MagneSphere® technology to eliminate the need for oligo(dT) cellulose and its associated problems. With total RNA as the starting material, the poly(A) mRNA fraction can be isolated from other nucleic acid contaminants in approximately 45 minutes. The isolated mRNA is suitable for all molecular biology applications, including in vitro translation and cDNA synthesis.
The system uses a biotinylated oligo(dT) primer to hybridize at high efficiency in solution to the 3´ poly(A) region present in most mature eukaryotic mRNA species. The hybrids are captured and washed at high stringency using streptavidin coupled to paramagnetic particles and a magnetic separation stand. The mRNA is eluted from the solid phase by the simple addition of ribonuclease-free deionized water. This procedure yields an essentially pure fraction of mature mRNA after only a single round of magnetic separation.