Recover RNA probes from denaturing acrylamide gels
The E.Z.N.A.® Poly-Gel RNA Extraction Kit is a straight forward and simple system for recovering RNA probes from denaturing acrylamide gels. After the in vitro transcription reaction, the product is run on a denaturing polyacrylamide gel (a mini protein gel apparatus can be used) to separate the DNA template, full-length RNA probe, any prematurely terminated products and free-nucleotides, by size. The gel is exposed to film and stained or UV shadowed (depending on the nature and quantity of the probe made, e.g. radioactive, non-isotopic or unlabeled probe). Full-length probe is identified and the band is cut from the gel. The probe is eluted by passive diffusion from the gel fragment and further purified using an HiBind® RNA spin-column. The procedure can produce enough probe, ready for hybridization in just 1-4 hours.