When you isolate mRNA using Dynabeads, you can also easily construct cDNA libraries for use in cDNA amplification, cDNA cloning, RACE experiments and subtractive hybridization.
When preparing solid-phase cDNA libraries, you first capture the mRNA onto the Dynabeads. The beads are then added directly to the RT-reaction, where the bead-bound oligo-dT residues act as primers for first strand cDNA synthesis. As all operations utilize the solid-phase, buffer changes are fast, simple and effective.
A stable, covalently linked cDNA library is produced and can be used over and over again (e.g for PCR-amplification, RACE, subtractive hybridisation, cDNA cloning etc.). Optimal buffer conditions are achieved by simply magnetic separation and resuspension in the appropriate buffer. Reverse transcription will only be limited by the efficiency of the enzyme used.
The bead-bound probe used both for capture and primer can be oligo-dT or any specific DNA sequence. The patented Dynabeads solid-phase cDNA synthesis technology is compatible with a various commercially available cDNA kits.
The use of Dynabeads Oligo(dT)25 for capture of mRNA for cDNA synthesis and/or subtractive cloning is covered by the patent "Process for producing" cDNA (WO9006044, EPO444119, AT122721T, DE68922743T, DE68922743D, CA2003500, AU627815, US5759820 and JP2960776B2).
For Research Use Only. Not for use in diagnostic procedures.