Poly(rA) Parallel Duplex Applications
We are developing a new method for isolating mRNAs from total RNA using the formation of poly(A) duplexes and analyzing our results using reverse transcriptase and RT-PCR amplification.
We are developing this method for the oligo(LNA-A) system because there is currently only one method, the traditional oligo(dT) method and we want to see if it can provide an alternative/better mRNA capturing method compared to the traditional approach. One of the goals is for the optimization of affinity oligo(LNA-A) purification of poly(rA)-tailed RNAs produced in testing buffer compositions, matrix material, blocking RNA, incubation time, and wash and elution procedure. Also, we are attempting to optimize and compare our approach to the oligo(dT) poly(rA) tailed mRNA transcript purification method. We anticipate low pH (e.g. 5.5) can be used in this method, which helps preserve the integrity of the RNAs better than the traditional oligo(dT) method that works at a pH of 7.2.