The use of Taka-diastase in a[3H]poly(A) hybridization assay of oligo(U) sequences in RNA.
A reliable assay of uridylate sequences longer than 10 is described. The procedure is based on the hybridization of [3H]poly(A) with poly(U) or oligo(U) sequences in high ionic conditions and a subsequent degradation of single stranded polynucleotides with purified Taka-diastase. A 1:2 complex between poly(A) and poly(U) is formed on which on poly(U) strand is digested by Taka-diastase. The procedure is especially suitable for the detection and quantitation of Un (n greater than 10) in RNA preparation.
D000681:Amylases / Q000378:metabolism; D016282:Aspartic Acid Endopeptidases; D001483:Base Sequence; D010450:Endopeptidases / Q000378:metabolism; D007700:Kinetics; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000032:analysis*; D009843:Oligoribonucleotides / Q000032:analysis*; D011061:Poly A; D012313:RNA; D014500:Uracil Nucleotides / Q000032:analysis*
D009841:Oligonucleotides; D009843:Oligoribonucleotides; D014500:Uracil Nucleotides; C027916:oligo(U); D011061:Poly A; D012313:RNA; D000681:Amylases; D010450:Endopeptidases; D016282:Aspartic Acid Endopeptidases; C022772:Aspergillus oryzae carboxyl proteinase