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用于核酸纯化的磁珠的最新进展。

Recent developments of magnetic beads for use in nucleic acid purification.

作者信息

Levison P R, Badger S E, Dennis J, Hathi P, Davies M J, Bruce I J, Schimkat D

机构信息

Whatman International Ltd., Maidstone, Kent, UK.

出版信息

J Chromatogr A. 1998 Aug 7;816(1):107-11. doi: 10.1016/s0021-9673(98)00064-8.

Abstract

The performance of Magarose, an agarose-based bead containing a paramagnetic component has been evaluated. The anion exchanger DEAE-Magarose is effective at binding DNA from a crude cell lysate. The plasmid pBluescript was isolated from 1.5 ml Escherichia coli JM109 cell culture, following alkaline lysis yielding 8.2 micrograms high-quality DNA. Under similar binding conditions 21 micrograms of salmon sperm DNA bound to the ion exchangers. The affinity medium oligo-dT Magarose was demonstrated to bind 75 mumol of an oligo-dA probe/g of medium by hybridization. Under similar conditions mRNA could be isolated from a preparation of baby hamster cell total RNA. The magnetic susceptibility of Magarose is very high, facilitating the use of this separation technique for rapid batch chromatographic processes.

摘要

对含有顺磁成分的琼脂糖基珠粒Magarose的性能进行了评估。阴离子交换剂DEAE - Magarose能有效地从粗细胞裂解物中结合DNA。从1.5毫升大肠杆菌JM109细胞培养物中分离出质粒pBluescript,经碱性裂解后得到8.2微克高质量DNA。在相似的结合条件下,21微克鲑鱼精DNA与离子交换剂结合。亲和介质寡聚dT Magarose通过杂交被证明每克介质能结合75微摩尔的寡聚dA探针。在相似条件下,可从小仓鼠细胞总RNA制剂中分离出mRNA。Magarose的磁化率非常高,便于将这种分离技术用于快速批量色谱过程。

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