Nucleic Acid Synthesis
NMR spectroscopy is an important tool to determine structures of nucleic acids alone and in interaction with proteins or small molecules. In fact, about half of the RNA structures deposited in the Protein Data Bank have been determined by NMR spectroscopy. Nucleic acids containing roughly less than ~40 nucleotides will require simple 15N or 13C15N enrichment to provide the constraints necessary to determine full three-dimensional structures. Advances in isotope-enrichment strategies have been crucial in the success for the study of larger RNAs. These advances primarily include the use of selectively and uniformly deuterated nucleotides and segmental isotope labeling using deuterium.
The most popular approaches to produce labeled RNA molecules for NMR studies use enzymatic in vitro transcription methods that employ labeled rNTPs, T7 RNA polymerase and either linearised plasmids or
double stranded DNA as templates. These techniques are used to construct labeled RNA molecules of which all of one type of nucleotide is labeled.
Labeled DNA oligonucleotides are routinely synthesized using enzymatic in vitro methods that utilize labeled dNTPs, a DNA polymerase, and a cDNA template. One advantage of using enzymatic methods over phosphoramidite chemistry is that large oligonucleotides (e.g., >50 nucleotides in length) can be prepared in milligram quantities. Position-specific labeled DNA molecules can be synthesised using standard phosphoramidite chemistry (using CIL’s deoxyphosphoramidites) to overcome the limited chemical-shift dispersion of DNA, as well as to create residue-specific probes to obtain functional, structural and dynamic information.
Please search our catalogue for any products of interest such as rNTPs, dNTPs, dNMPs, deoxyphosphoramidites etc.
Below are links to our Structural Biomolecular NMR catalogue and a couple of relevant application notes written by CIL customers.
If you have any questions or wish to discuss your needs then please don’t hesitate to contact us.
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