General information on PNA synthesis and purification

Synthesis
PNA synthesis is carried out by FMOC chemistry under continuous flow conditions using Rink Amide PEGA resins, which means that the C terminal is automatically amidated. At the completion of synthesis, PNA are cleaved from the resin and de-protected using TFA/m-cresol cleavage cocktail. PNA are then precipitated from the cleavage cocktail using cold ether after remove TFA. The precipitate is then washed 2~3 times with the chilly ether and then dissolved in a crude solution containing 0.1% of TFA prior to purification or lyophilization.

Purification:
Successful synthesis is dependent on a lot of factors, especially the sequence of the PNA and the coupling efficiency of PNA monomer in the synthesis. The ultimate purity of the synthesized product is also related to the sequence. Impurities such as salts, deletion PNAs (created due to incomplete coupling), protecting groups and modified PNAs can vary from 10% to 90% depending on PNA sequences. It is recommended that all the PNAs should be purified prior to use. Purification synthetic PNA is carried out on a semi-preparative HPLC system using a RP C-18 column and gradient elution with 5~40% mixture of solvent A (0.1% of TFA in water) and B (0.1% of TFA in CAN). Fractions containing the PNA will be collected, QC by MALDI & HPLC, and lyophilized for shipping.

Notes:
PNA QC is based on an analytical-scale reverse-phase HPLC chromatogram and the mass analysis by MALDI. It will be provided for every PNA to ensure the highest quality of synthetic product.

Research PNA synthesis is licenced by Panagene. Clinical and commercial use please contact life technology and Isis