Adsorption chromatography of nucleic acids on silicone-coated porous glass.
Bovine liver tRNA was adsorbed on silicone-coated porous glass in 5 M NaCl, 10 mM Tris-HCl (pH 7.6) and fractionated by elution with decreasing NaCl concentrations. tRNAPro, tRNAVal, tRNAIle, tRNAThr, tRNASer, and tRNAPhe were eluted in this order. tRNA which had been digested with ribonuclease A was not adsorbed. Q beta RNA (adsorbed onto the glass in 5 M NaCl) was eluted with 1.5 M NaCl. RNA species in a crude rRNA fraction from Escherichia coli were separated into tRNA, 5S rRNA, and high molecular weight rRNA on siliconized porous glass. A half of calf thymus DNA was adsorbed on the glass in 5 M NaCl and the residual part passed through the column. The CD spectra showed that DNA and tRNA took the C-form and the A-form in 5 M NaCl, respectively. Therefore, the discrepancies of behavior of the DNA and RNA on siliconized porous glass may be related to the occurrence of these forms. The recovery of these nucleic acids from the column was 83-100%. Adsorption chromatography on siliconized porous glass may be a useful method for the separation of tRNA, rRNA, and mRNA.
D000818:Animals; D002417:Cattle; D002845:Chromatography / Q000379:methods; D005898:Glass; D008099:Liver / Q000032:analysis; D008970:Molecular Weight; D009696:Nucleic Acids / Q000302:isolation & purification*; D012343:RNA, Transfer / Q000302:isolation & purification; D012346:RNA, Transfer, Amino Acyl / Q000302:isolation & purification; D012828:Silicones; D013950:Thymus Gland / Q000032:analysis
D009696:Nucleic Acids; D012346:RNA, Transfer, Amino Acyl; D012828:Silicones; D012343:RNA, Transfer