The invention pertains to a three step initiation, elongation and termination method and platform for synthesizing selectively labeled RNA molecules by first polymerizing a first liquid phase RNA molecule from a solid phased DNA template fixed onto a solid phase.
Background:
The method includes the steps of incubating the solid and liquid phases at appropriate elongation temperatures and then terminating elongation by a separation stage where the phases are incubated at near 0 degrees Celsius where it selectively terminates RNA elongation. The steps can be repeated by the number bases (rNTPs) in the final RNA molecule wherein in each iterative stage a new rNTP can be added that is selectively labeled. The DNA may have a density of 30-80% on the solid substrate, and the solid substrate may be a bead. The bead may comprise a gel, glass, or a synthetic polymer. The bead may have a diameter of 5-100 mm. The concentration of DNA may be 30 mm-1 nm. The concentration of rNTP may be 1-100 times the DNA concentration. The RNA polymerase may be a T7 RNA polymerase. The label may be 13 C/5 N, 2 H, Cy3, Cy5, a fluorophore, a heavy atom, or a chemical modification.