Magnetic DNA hybridization properties of oligonucleotide probes attached to superparamagnetic beads and their use in the isolation of poly(A) mRNA from eukaryotic cells

Erik Hornes, Lars Korsnes
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引用次数: 47

Abstract

Poly(A) messenger RNA is generally purified from total RNA using oligo(dT) cellulose affinity chromatography or centrifugation through spin columns. We present a new method for rapid purification of poly(A) mRNA using oligo(dT) probes attached to superparamagnetic beads. By magnetic separation, washing, and elution, pure mRNA is obtained from living cells within 10 minutes. This procedure works for crude RNA preparations or cell lysates that would otherwise clog standard oligo(dT) cellulose column systems. The present method reduces the risk of degradation, is highly efficient, and can easily be scaled up or down.

超顺磁珠寡核苷酸探针的磁DNA杂交特性及其在真核细胞中聚(A) mRNA分离中的应用
聚(A)信使RNA通常是从总RNA中纯化使用寡聚(dT)纤维素亲和层析或离心通过自旋柱。我们提出了一种快速纯化poly(a) mRNA的新方法,使用附着在超顺磁珠上的oligo(dT)探针。通过磁分离、洗涤和洗脱,在10分钟内从活细胞中获得纯mRNA。该程序适用于粗RNA制剂或细胞裂解物,否则会堵塞标准寡聚(dT)纤维素柱系统。本方法降低了降解的风险,效率高,并且可以很容易地按比例放大或缩小。
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