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preQ1-biotin is a modified analog of the natural substrate prequeosine1 (preQ1) and is used for RNA-TAG (transglycosylation at guanosine) and DNA-TAG. preQ1-biotin enables affinity tagging and pull-down of specific RNAs that have been modified selectively by E. coli tRNA guanine transglycosylase (TGT). preQ1-biotin is incorporated site-specifically and covalently into RNAs containing a short harpin nucleotide recognition motif. The guanine in a UGU recognition element is exchanged with the preQ1-biotin substrate.
Plasmids for expression of the E.coli TGT enzyme (#138201) and for cloning an RNA of interest into a vector containing the recognition motif at the 3' end (#138209) are available from Addgene.
E. coli TGT irreversibly exchanges a specific guanine in a DNA hairpin with preQ1-biotin. Schematic representation of preQ1-bitoin insertion into DNA.
E. coli TGT irreversibly exchanges a specific guanine in a DNA hairpin with preQ1-biotin. Urea-PAGE gel shift analysis of preQ1-biotin insertion. Modification is evidenced by an upward shift of the product band due to molecular weight increase. Lane 1 is unmodified DNA; lane 2 is preQ1-biotin modified DNA.
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|Storage||Store at -20°C|
The technical data provided above is for guidance only. For batch specific data refer to the Certificate of Analysis.
Tocris products are intended for laboratory research use only, unless stated otherwise.
|Solvent||Max Conc. mg/mL||Max Conc. mM|
The following data is based on the product molecular weight 504.65. Batch specific molecular weights may vary from batch to batch due to the degree of hydration, which will affect the solvent volumes required to prepare stock solutions.
|Concentration / Solvent Volume / Mass||1 mg||5 mg||10 mg|
|0.2 mM||9.91 mL||49.54 mL||99.08 mL|
|1 mM||1.98 mL||9.91 mL||19.82 mL|
|2 mM||0.99 mL||4.95 mL||9.91 mL|
|10 mM||0.2 mL||0.99 mL||1.98 mL|
References are publications that support the biological activity of the product.
Busby et al (2020) Enzymatic RNA biotinylation for affinity purification and identification of RNA-protein interactions. ACS Chem.Biol. 15 2247 PMID: 32706237
Tota and Devaraj (2023) Site-specific covalent labeling of DNA substrates by an RNA transglycosylase. J.Am.Chem.Soc 145 8099 PMID: 36988146
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Keywords: preQ1-biotin, preQ1-biotin supplier, preq1biotin, prequeosine1, RNA, binding, labeling, purification, TGT, rna, tag, rnatag, rna-tag, dna-tag, dnatag, Biochemicals, and, Molecular, Biology, 7804, Tocris Bioscience
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