Capillary electrophoresis of DNA with high resolution based on copoly(pentaerythritoltetra succinimidylcarboxypentyl/aminopropyl polyoxyethylene) hydrogel

Capillary gel electrophoresis is widely applied for determination of sequence and size of DNA, in which the sieving gel plays an unignorable role. Herein, a pore-size controllable hydrogel was synthesized in the capillary with two symmetrical tetrahedron-like macromonomers consisting of pentaerythritoltetra (succinimidylcarboxypentyl) polyoxyethylene (PS) and pentaerythritoltetra (aminopropyl) polyoxyethylene) (PA). By capillary electrophoresis of the DNA fragments with this hydrogel, it is found that a homogenous structure of hydrogel which is more suitable for the DNA separation can be achieved when the molecular weight of PA is approximate to that of PS. DNA fragments smaller than 1500 bp can be well resolved in this hydrogel within 13 min. More than 100 consecutive runs can be carried out in such a dynamically coated capillary before performance begins to degrade. Notably, such hydrogel can realize separation of dsDNA up to single base pair resolution and same length of dsDNA with 1 bp difference. (c) 2021 Elsevier B.V. All rights reserved.

Automated summary

Capillary gel electrophoresis using a controllable pore-size hydrogel can achieve a homogenous structure suitable for DNA separation based on the molecular weights of PA and PS. This hydrogel allows for efficient resolution of DNA fragments smaller than 1500 bp and more than 100 consecutive runs before performance degradation. Notably, the hydrogel can separate dsDNA up to single base pair resolution and differentiate dsDNA of the same length with 1 bp difference.