普通级琼脂糖及低电内渗琼脂糖制备工艺

Preparation Process of Regular Agarose and Low Electroendosmosis Agarose

以琼脂粉为原料,采用磷酸氢二钠-柠檬酸缓冲液法制得普通级琼脂糖。选取琼脂粉添加量为20 g/L、提取温度为40℃、提取时间为6 h的工艺条件进行放大试验,分别进行琼脂粉20、60、900 g试验,其制得的琼脂糖电内渗值分别为0.22、0.20、0.18,得率分别为88.50%、89.20%、85.12%,凝胶强度分别为850、876、865 g/cm2。进一步采用DEAE-纤维素树脂层析法将普通级琼脂糖精制为低电内渗琼脂糖,其电内渗值为0.065,但凝胶强度不足。按1∶1将普通级琼脂糖与低电内渗琼脂糖混合后的样品,其电内渗值为0.11,仍然符合低电内渗指标的要求,同时其凝胶强度大大增强,可以进行电泳操作。

Regular agarose was prepared from agar using the method of disodium hydrogen phosphate-citric acid buffer. In amplification experiment with the parameters which were the quantity of agar 20 g/L, extraction temperature 40℃ and extraction time 6 h, the product yield of the agarose sample prepared from 20 g, 60 g and 900 g agar was 88.50%, 89.20% and 85.12%, respectively. The electroendosmosis （EEO）of the agarose sample prepared from 20 g, 60 g and 900 g agar was 0.22,0.20 and 0.18, respectively. The gel strength （GS）of the agarose sample prepared from 20 g, 60 g and 900 g agar was 850 g/cm^2, 876 g/cm^2 and 865 g/cm^2,respectively. The regular agarose sample was further refined to be low electroendosmosis agarose sample by the method of DEAE-Cellulose. The results showed that EEO of the prepared agarose sample were 0.065 ,but the GS was lower. After mixing regular agarose sample with low electroendosmosis agarose sample in 1：1, the EEO of the mixed sample was 0.11,which met requirements for low electroendosmosis, at the same time, the GS greatly enhanced to be suitable for gel electrophoresis analysis.