PNA synthesis is usually performed by automated solid phase methods, as in the case of DNA and RNA oligonucleotide synthesis. BOC Sciences provides professional PNA synthesis services, in addition to routine synthesis, as well as professional peptide nucleic acid-related technical support to assist customers in their research and development efforts.
Fig. 1. General protocol for the synthesis of PNA. (Elskens et al., 2017)
PNA is a nucleic acid analog in which the normal phosphate bonds in DNA and RNA are replaced by a neutral peptide-like N-(2-aminoethyl)glycine backbone. Different basic groups (purines and pyrimidines) are then attached to the peptide framework via a methylene carbonyl group to form a non-chiral and uncharged structure to ensure chemical stability against hydrolysis reactions. Since PNA has no charged phosphate groups, there is no electrostatic repulsion. This makes the binding of PNA to DNA stronger than that between DNA and DNA. PNA is neither a peptide nor a nucleic acid, so neither DNA enzymes nor proteases can hydrolyze PNA, and PNA is quite stable in vitro and in vivo.
BOC Sciences requires PNA design prior to PNA synthesis, and these designs can be performed according to customer requirements. After the PNA design is completed, the peptide nucleic acid is then custom synthesized.
Due to the high affinity of PNAs, it is not necessary to design very long sequences, generally 12-18 is sufficient. For PNA, sometimes shorter sequences will also achieve the desired results.
PNAs with high purine content, especially guanine G, are prone to aggregation precipitation. Therefore, do not have 6 or more purines present in any 10 consecutive PNA monomers.
When the PNA chain is long (>22mer) or the purine content is high (>60%), it is recommended to include some solubilizing groups or two lysines in the sequence in order to improve the solubility of the PNA probe. When PNA is coupled to a peptide or dye, the linker can act as a spacer.
In PNA synthesis, the carboxylic acid is first activated with a benzotriazole urea salt, which is then combined with HOBt and diimide to produce a hydroxybenzotriazole ester. At the end of each PNA synthesis cycle, the PNA oligonucleotides were treated with a DMF solution containing 20% piperidine in order to remove the Fmoc protecting group.
BOC Sciences offers fluorescence modification, biotin modification, peptide modification, and glycosylation modification of PNA.
Routine synthesis scale: 50, 100, 1000 nmole, free synthesis report including HPLC and MS profiles for purity and structure characterization.
If you are interested in our services, or your needs are not in the list for the time being, please feel free to contact us, we will customize an experimental solution for you.