Nucleotide separation

Example 45 Mizuguchi and Makino have observed stereoselective phos-phitylation of thymidine by P-aryloxy nucleotide separated into diastere-omers [80]. 

Fig. 2.11. Thin-layer chromatogram and corresponding peaks for some nucleotides separated on purified cellulose. The arrows indicate the direction of the reflectance scan.

Advances in enzymatic digestion methodology and nucleotide separation have made it possible to study the primary structures of the messenger RNAs — 3000-4000 residues) as well as of DNAs (10 -10 residues). 

Na,K-ATPase activity. The hormone response element for Na,K ATPase or, more accurately one of its subunits, is AGGTCACTCCGGGACG Feng et ol., 1993). The half-core se juences are underlined. Note that four nucleotides separate the halfcore sequences, and that one of these sequences deviates from the typical AG-GTCA sequence. Thyroid hormone s stimulation of mitochondrial proteins and Na,K-ATPase is consistent with this hormone s action in raising the body s metabolic rate. 

Many schemes for fractionating nucleotides, nucleosides and bases on sulphonated polystyrene resins have been published. The main difficulty with these methods is variation between resin batches (e.g. Anderson et al. 1963). Nucleotide separations can be achieved on DEAE-cellulose (Whatman Data Sheet 13, 1967) and DEAE-Sephadex (Piers et al. 1965b) but these media do not seem to be widely used. Gel filtration columns will separate some nucleotide components. Ligand exchange chromatography and partition chromatography of nucleosides are useful for minor components. 

Some ribonucleoside and nucleotide separations on 1.5x34 cm columns of DEAE-cellulose eluted with 0.09 M sodium acetate, 0.2 M acetic acid pH 4.4 have been described (Whatman Data Sheet DS/13). 

The selectivity of ion-exchange chromatography for nucleotide separations is significantly affected by the pH of the mobile phase and this can therefore be manipulated to achieve many separations. 

Target Oligo- nucleotide Separation System Detection System Model Ref. 

In 1988, it was shown that DNA single nucleotide separation could be achieved using cross-linked polyacrylamide gel filled capillaries coupled with U V/Vis detection [7]. These chemical gels provided... 

FIGURE 3.17 IM-MS 2D plot of a short oligonucleotide, apeptide, and a lipid. Note the various trend lines along which these compounds and their fragments are aligned. (From Woods et al., Lipid/peptide/nucleotide separation with MALDI-ion mohility-TOF MS, Anal. Chem. 2004, 76(8), 2187-2195. With permission.)... 

Woods, A.S. Ugarov, M. Egan, T. Koomen, J. Gillig, K.J. Fuhrer, K. Gonin, M. Schultz, J.A., Lipid/peptide/nucleotide separation with MALDl-ion mobihty-TOF MS,... 

TLC has been combined with electrophoresis to facilitate nucleotide separations. Thus, Berquist (1965) mapped the enzymatic digests of RNA by separating 17 ohgonucleotides and mononucleotides on a cellulose layer. Electrophoresis was first run at pH 2.5 and 50 V/cm, followed by two successive chromatographic developments at a right angle to the electrophoretic dimension. Diels and Wachter (1980) and Hashimoto et al. (1981) also combined gel electrophoresis and TLC in their RNA sequencing studies. 

Woods AS, Fuhrer K, Egan T, Ugarov M, Koomen J, Gonin M, GiUig KJ, and Schultz JA. Lipid/Peptide/Nucleotide Separation with MALDI-Ion Mobdity-TOFMS. Anal. Chem. 76, 2187-2195 (2004). 

Nucleotides Separation. A method for the separation of nucleotides has been developed (10). These nucleotides include 5 -monophosphate adenosine, 5 -monophosphate cytidine, 5 -monophosphate uridine, 5 -monophosphate guanosine, and... 

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