practical-aspects-sample-preparation-setup-1h-r1-relaxation

Practical Aspects of Sample Preparation and Setup of 1H R1&#961; Relaxation Dispersion Experiments of RNA

RNA is a highly flexible biomolecule, wherein changes in structures play crucial roles in the functions that RNA molecules execute as cellular messengers ...

Karolinska Institutet

Pulse sequences in NMR spectroscopy sometimes require the application of pulses with effective flip angles different from 90° and 180°. Previously (Magn. Reson. Chem. 2015, 53, 886-893), offset-compensated broadband excitation pulses with RF-amplitude-dependent effective flip angles (RADFA) were introduced that are applicable in such cases. However, especially RF-amplitude-restricted RADFA pulses turned out to perform not as good as desired in terms of achievable bandwidths. Here, a class of RF-amplitude-restricted RADFA pulses with linear phase slope is introduced that allows excitation over much larger bandwidths with better performance. In this theoretical work, the basic principle of the pulse class is explained, their physical limits explored, and their properties, also compared with other pulse classes, discussed in detail. Copyright © 2017 John Wiley & Sons, Ltd.

Broadband RF-amplitude-dependent flip angle pulses with linear phase slope.

There is an increasing demand for efficient and robust production of short RNA molecules in both pharmaceutics and research. A standard method is  transcription by T7 RNA polymerase. This method is sequence-dependent on efficiency and is limited to products longer than ~12 nucleotides. Additionally, the native initiation sequence is required to achieve high yields, putting a strain on sequence variability. Deviations from this sequence can lead to side products, requiring laborious purification, further decreasing yield. We here present transcribing tandem repeats of the target RNA sequence followed by site-specific cleavage to obtain RNA in high purity and yield. This approach makes use of a plasmid DNA template and RNase H-directed cleavage of the transcript. The method is simpler and faster than previous protocols, as it can be performed as one pot synthesis and provides at the same time higher yields of RNA.

Hannes Feyrer

Department of Medical Biochemistry and Biophysics

Karolinska Institutet

Practical Aspects of Sample Preparation and Setup of ¹H R_1ρ Relaxation Dispersion Experiments of RNA

Hannes Feyrer

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Medical Biochemistry and Biophysics

Karolinska Institutet

Practical Aspects of Sample Preparation and Setup of 1H R1ρ Relaxation Dispersion Experiments of RNA

Department of Medical Biochemistry and Biophysics

Practical Aspects of Sample Preparation and Setup of ¹H R_1ρ Relaxation Dispersion Experiments of RNA