Microliter volumes of reactions were fractionated on EMD-Millipore thin-layer PEI-cellulose F eluted with 0.5 M LiCl at room temperature (Randerath 1968 ), then dried and phosphorimaged (BioRad FX). The characteristic positions shown in Figure 1 for 2′-5′, 3′-5′, and 5′-5′ A dinucleotides were identified using standard compounds, nuclease digestions, and mass spectrometry on eluted spots from reactions.
Emd millipore thin layer pei cellulose f
Manufactured by Merck Group
EMD-Millipore thin-layer PEI-cellulose F is a type of thin-layer chromatography (TLC) plate. It is composed of polyethyleneimine (PEI) coated on a cellulose matrix. The PEI coating provides a cationic exchange surface for the separation and analysis of various compounds.
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2 protocols using emd millipore thin layer pei cellulose f
1
Separation and Identification of A Dinucleotides
2
TLC Analysis of Radiolabeled Dinucleotides
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Samples were pipetted onto the origin on EMD-Millipore thin-layer PEI-cellulose F which had been eluted with deionized water and dried. Sheets are subsequently eluted at room temperature with 0.5 M LiCl (Randerath 1968 ). Chromatograms were dried and phosphorimaged (BioRad FX). [32P]AppA concentrations are calculated by multiplying the fraction of total cpm in the product spot by the known total concentration of AMP. Characteristic mobilities in Figure 1 for 2′–5′, 3′–5′, and 5′–5′ A dinucleotides were determined using characterized standards.
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