PURIFICATION OF MRNA BY SPECIFIC REMOVAL OF RRNA

THIS RESEARCH PROJECT WILL PROVIDE A NEW AND IMPROVED WAY IN WHICH RESEARCHERS CAN TREAT PREPARATIONS OF TOTAL RIBONUCLEIC ACID (RNA) TO OBTAIN MESSENGER RNA (MRNA), FREE OF RIBOSOMAL RNS (RRNA). THE TRADITIONAL METHOD HAS BEEN TO CAPTURE THE MRNA, WHICH HAS LONG REPEATS OF ADENINES (A) AT ITS 3' END, BY USING IMMOBILIZED OLIGO (DT) (16-30 THYMIDINES LONGS), AND THEN WASH AWAY THE RRNA WHICH DOES NOT BIND. THIS WORK WILL PROVIDE A WAY TO REMOVE RRNA FROM RNA PREPARATIONS, LEAVING THE MRNA. THIS WILL BE DONE BY PASSING THE RNA PREPARATIONS OVER BEADS THAT HAVE OLIGONUCLEOTIDES (THESE ARE SHORT SINGLE STRANDED DEOXYNUCLEOTIDE SEQUENCES) ATTACHED TO THEM WHICH ARE COMPLEMENTARY TO CONSERVED SEQUENCE OF RRNA. IN THIS WAY THE RRNA WILL BE BOUND AND MRNA WILL FLOW THROUGH. THIS NEW METHOD WILL BE ATTRACTIVE TO RESEACHERS AND THUS HAVE COMMERCIAL APPLICATION BECAUSE IT WILL PROVIDE A BETTER WAY TO OBTAIN MRNA. FIRST, THE YIELDS OF MRNA SHOULD BE HIGHER SINCE EVEN MRNA WITH SHORT POLY A TAILS WILL BE ISOLATED. SECONDLY, IT GIVES RESEARCHERS ACCESS TO NONPOLYADENYLATED MRNA'S WHICH ARE LOST IN THE OLIGO (DT) COLUMN METHOD.