Rapid preparation of RNA samples using DNA-affinity chromatography and DNAzyme methods.
Milligram quantities of RNA are commonly synthesized by in vitro transcription from a DNA template with T7 RNA polymerase. However, the run-off transcription method results in heterogeneity at the RNA 3'-terminus. RNA purification requires single-nucleotide resolution to separate the transcript of the correct length from the aborted or add-on transcripts that are usually present in comparable amounts. Here, we describe an RNA preparation method that uses a trans-acting DNAzyme and sequence-specific affinity column chromatography. This purification method is simple, fast, and suited for high throughput.
Division of Magnetic Resonance Research, Korea Basic Science Institute, Ochang, Cheongwon, South Korea.
Hae-Kap
HK
Cheong
Eunha
E
Hwang
Chaejoon
C
Cheong
D002846:Chromatography, Affinity / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D021881:DNA, Catalytic / Q000378:metabolism*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification*; D059367:RNA Cleavage; D013997:Time Factors; D014158:Transcription, Genetic
D021881:DNA, Catalytic; D012313:RNA; D004247:DNA