Polyethylenimine cellulose thin layer chromatography plates
Manufactured by Macherey-Nagel
Polyethylenimine cellulose thin-layer chromatography plates are a type of chromatographic media manufactured by Macherey-Nagel. They are used for the separation and analysis of various chemical compounds through thin-layer chromatography techniques.
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Lab products found in correlation
Vaccinia virus capping enzyme, by New England Biolabs (2 mentions)
Fluorescent image analyzer fla 3000, by Fujifilm (2 mentions)
Nuclease p1, by Fujifilm (2 mentions)
α 32p gtp, by PerkinElmer (2 mentions)
Glycogen, by Thermo Fisher Scientific (1 mentions)
Strataclean beads, by Agilent Technologies (1 mentions)
G 25 column, by GE Healthcare (1 mentions)
Nuclease p1, by Merck Group (1 mentions)
2 protocols using polyethylenimine cellulose thin layer chromatography plates
1
Enzymatic Synthesis and Analysis of Radiolabelled RNA Caps
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G *pppGGGACAAGU (in which the asterisk indicates the [32P]-labelled phosphate) was synthesized by incubating pppGGGACAAGU (10 μM) with vaccinia virus capping enzyme (New England Biolabs) in the presence of 1.65 μCi [α-32P]-GTP (Perkin Elmer). The capped RNA was purified by precipitation in 3 M sodium acetate supplemented with 1 μg μl−1 of glycogen (Thermo Scientific), and submitted to methylation by CR-VI+ (as above), after which it was precipitated again (stopping the reactions), and digested with 1 U of Nuclease P1 (US Biologicals) in 30 mM sodium acetate (pH 5.3), 5 mM ZnCl2 and 50 mM NaCl (4 h, 37 °C). The products were spotted onto polyethylenimine cellulose thin-layer chromatography plates (Macherey Nagel), and resolved in two steps, first using 0.65 M LiCl, then 0.45 M (NH2)2SO4 as mobile phase. The radiolabelled caps released by Nuclease P1 were visualized using a Fluorescent Image Analyzer FLA3000 (Fuji) phosphor-imager.
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2
Radioactive Capped RNA Synthesis
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Radioactive capped RNAs (G*pppRNA) were synthesized by incubating pppRNA (10 μM) with vaccinia virus capping enzyme (New England Biolabs) in the presence of 1.65 mCi of [α-32P]-GTP (Perkin Elmer). Labeled RNAs were purified with StrataClean beads (Agilent) to remove proteins and G25 columns (GE Healthcare) to remove excess of radioactive GTP. RNA was then submitted to methylation by the SUDV MTase+CTD domain (as above), and precipitated as described for the in vitro transcription products. Finally, RNAs were digested with 1 U of nuclease P1 (Sigma) in 30 mM sodium acetate (pH 5.3), 5 mM ZnCl2 and 50 mM NaCl (4 h, 37°C). Products were spotted onto polyethylenimine cellulose thin-layer chromatography plates (Macherey-Nagel) and resolved using 0.65 M LiCl as mobile phase. The radiolabeled caps released by nuclease P1 were visualized using a Fluorescent Image Analyzer FLA3000 (Fuji) phosphor-imager.
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