Preparation of Agarose from Gelidium amansii for Gel Electrophoresis using Various Purification Methods and Its Resolution Characteristics for DNA

다양한 정제방법에 의한 전기영동용 한천유래 아가로즈의 제조 및 DNA분리 특성

The present study was conducted to investigate the preparative methods of agarose for gel electrophoresis from agar. Naturally occuring agar consists of two main polysaccharides, the neutral polysaccharide agarose and the acid sulphated polysaccharide agaropectin. The sulphate and carboxyl functions of the agar are accumulated in the agaropectin. The hydrophilic, non-ionogenic, rigid and transparent gel matrix of the agarose was found to be suitable for gel electrophoresis gel filtration and affinity chromatography. Agar was purified by chitosan treatment, cetylpyridinium chloride (CPC) treatment, and polyethylene glycol (PEG) treatment. Yields of agarose purified from agar with chitosan, CPC and PEG were 56.7%, 55.6% and 62.3%. It was proper to treat with chitosan in preparative methods of agarose for gel electrophoresis from agar.

제주도산 우뭇가사리로부터 agar를 제조하여 분석용 등급의 agarose 제조를 위하여 cetylpyridium chloride (CPC), polyethylene glycol (PEG), chitosan을 처리하여 agaropectin을 제거하여 agarose를 분리, 정제하였다. 3가지 방법중 chitosan을 처리하여 정제한 agarose는 황산기 함량이 가장 낮았으며, electroendosomosis도 가장 낮은 수치를 나타내어 분석용 agarose 제조를 위한 가장 효율적인 방법으로 나타났으며 실제 DNA 전기영동 실험에서도 가장 우수한 resolution을 나타내었으며 교차면역 전기영동에서도 시판되고 있는 제품에 비하여 품질이 양호함을 알 수 있었다.