Poly(U)-agarose affinity chromatography: specific, sensitivity selectivity, and affinity of binding.
These studies were done to determine four basic intrinsic properties of poly(U)-agarose affinity columns. Specificity of binding studies demonstrated that binding to these columns is highly specific with greater than 90% complementary binding and less than or equal to 3% noncomplementary binding. Sensitivity of binding studies indicated that a minimum sequence of 10 adenylates is required for detectable complementary binding. Selectivity of binding studies revealed that nonsequential adenylates in native RNAs and randomly distributed adenylates in synthetic poly(A)-poly(C) co-polymers did not bind to poly(U)-agarose affinity columns. Whereas, affinity of binding studies demonstrated that A=U complementary base pairing is independent of chain-lengths of greater than or equal to 25 adenylates and dependent of chain-lengths of less than 25 adenylates. Thus the data demonstrates that poly(U)-agarose affinity chromatography is scientifically sound and expedient for the detection and isolation of poly(A)-containing cellular and viral RNAs.
D002846:Chromatography, Affinity / Q000379:methods*; D004926:Escherichia coli; D009843:Oligoribonucleotides / Q000302:isolation & purification; D011061:Poly A / Q000302:isolation & purification; D011072:Poly U; D011131:Polyribonucleotides / Q000302:isolation & purification*; D012329:RNA, Bacterial / Q000302:isolation & purification*; D012343:RNA, Transfer / Q000302:isolation & purification*; D012685:Sepharose
D009843:Oligoribonucleotides; D011131:Polyribonucleotides; D012329:RNA, Bacterial; D011061:Poly A; D011072:Poly U; D012685:Sepharose; D012343:RNA, Transfer