Terms of chemistry and functionality, 2′-OMe has been used in a much broader range of applications, some of which are unrelated to therapeutics. These include aptamers,6 detection probes,7 RNAi,8 DNAzymes/ribozymes9, 10 and CRISPR.11 2′-MOE should be applicable in all these contexts, as it has superior duplex stability and nuclease resistance. We hope that the addition of 2′-MOE will enable researchers to expand in their use of it. The use of 2′-MOE reagents in oligonucleotide synthesis is relatively straightforward. A coupling time of 6 min is recommended, and oligonucleotides that contain these residues can be deprotected following our standard procedures. It is important to note that methylamine should not be used with 2′-MOE-Bz-5-Me-C, in order to avoid methylation of the N4.
Application Note — Trimer Phosphoramidites
Glen Research offers a unique line of products known as trimer phosphoramidites. These are trinucleotide reagents that allow customers to effectively synthesize oligonucleotides based on amino acid codons rather than individual nucleotides.1 For researchers looking to generate oligonucleotide libraries for mutagenesis, these reagents avoid stop codons and amino acid redundancy.72795-01-8 SMILES In theory, such libraries can be obtained via a split and pool synthesis workflow without trimer phosphoramidites, but this is very awkward to perform in practice for a large number of codons or longer codoncontaining regions. Glen Research offers a total of 29 trimer phosphoramidites that cover all 20 standard amino acid codons in the sense and antisense directions and are optimized for E. coli expression. We also offer two standard trimer phosphoramidite mixes, both for sense trimers: Mix 2, 13-1992xx, which contains an equal mixture of 19 codons (no cys), and Mix 1, 13-1991-xx, which contains an equal mixture of 20 codons. For other ratios, we can custom prepare ready-to-use reagents as desired. Alternatively, customers may purchase the trimers individually and then construct their own mixes. Those who do decide to pursue this latter option will need to be mindful of the different coupling rates that the various trimers exhibit, as detailed in Glen Report articles 16.25 and 25.12. Over the years, our customers have published some exciting results using libraries synthesized with our trimer phosphoramidite mixes. The Regan Group at Yale developed a new screening method for protein affinity agents.2 The technique is based on splitting green fluorescent protein (GFP) into 2 halves and fusing a peptide library and a target peptide to the GFP fragments (Figure 1). During screening, if a member of a library binds to
Figure 1.1599432-08-2 web Split-GFP reassembly assay the desired peptide, then the two halves of GFP are brought together resulting in detectable fluorescence.PMID:25905391 The researchers constructed a peptide library based on the tetratricopeptide repeat (TPR) as a framework (34 amino acids, aa). In the library, seven positions were randomized with our Trimer Phosphoramidite Mix 2 (no cys). This library was fused with the C-terminal fragment of GFP (CGFP) and screened against several targets in E. coli: c-Myc epitope tag (10 aa), full length Dss1 (70 aa), and the C-terminal epitope of Dss1 (19 aa), each of which was fused to the N-terminal fragment of GFP (NGFP). Screening was performed in E. coli, and bacteria that were fluorescent were identified and isolated by fluorescence activated cell sorting (FACS). For each of the targets, two rounds of iterative selecti.MedChemExpress (MCE) offers a wide range of high-quality research chemicals and biochemicals (novel life-science reagents, reference compounds and natural compounds) for scientific use. We have professionally experienced and friendly staff to meet your needs. We are a competent and trustworthy partner for your research and scientific projects.Related websites: https://www.medchemexpress.com
