A simple and efficient method to transcribe RNAs with reduced 3' heterogeneity.
Ribose C2' methoxy groups (-OCH3) on the penultimate nucleotide or the last two nucleotides of DNA templates can dramatically improve the quality of transcripts produced by T7 RNA polymerase. This strategy can be adapted to generate transcripts of varying lengths and will allow greater ease in purification of the products.
Kao
C
C
Department of Biology, Indiana University, Bloomington, Indiana 47405, USA. ckao@bio.indiana.edu
Rüdisser
S
S
Zheng
M
M
D004247:DNA / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D005821:Genetic Techniques; D012313:RNA / Q000235:genetics*; D012266:Ribose / Q000737:chemistry; D014158:Transcription, Genetic; D014764:Viral Proteins
D014764:Viral Proteins; D012313:RNA; D012266:Ribose; D004247:DNA; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases