Analysis of double-stranded poly(A).poly(U) molecules by reversed-phase high-performance liquid chromatography.
The behaviour of different batches of synthetic Poly(A).Poly(U) in reversed-phase high-performance liquid chromatography (HPLC) was studied. They consist of large molecules mainly in the form of a double strand. Differences in the elution patterns were correlated with properties detected by conventional methods such as electrophoresis, centrifugation, fusion analysis or enzymatic digestions. Under the present conditions, contamination by products and precursors used during synthesis was detectable, but was absent in most of the preparations. The differences in elution patterns between batches appear to be correlated with the size of the molecules synthesized. The chromatograms suggested that Poly(A).Poly(U) molecules contain single-strand portions at least transiently. The presence of such portions was confirmed by enzymatic digestion with S1 nuclease. The rapidity, reproducibility and ease of reversed-phase HPLC qualify this technique as a tool for routine analysis.
Liautard
J
J
U-249 INSERM, CRBM du CNRS, Université Montpellier I, France.
Colote
S
S
Ferraz
C
C
Sri-Widada
J
J
Pichot
A
A
Liautard
J P
JP
D002851:Chromatography, High Pressure Liquid; D004587:Electrophoresis, Agar Gel; D011063:Poly A-U / Q000032:analysis*; D013056:Spectrophotometry, Ultraviolet
D011063:Poly A-U