Hornes E, Korsnes L
Apothekernes Laboratorium A.S., Oslo, Norway.
Genet Anal Tech Appl. 1990 Oct;7(6):145-50. doi: 10.1016/0735-0651(90)90028-e.
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.
聚腺苷酸信使核糖核酸(Poly(A) mRNA)通常使用寡聚(dT)纤维素亲和色谱法或通过旋转柱离心从总核糖核酸(RNA)中纯化出来。我们提出了一种使用附着在超顺磁性珠上的寡聚(dT)探针快速纯化聚腺苷酸信使核糖核酸(Poly(A) mRNA)的新方法。通过磁分离、洗涤和洗脱,可在10分钟内从活细胞中获得纯信使核糖核酸(mRNA)。该程序适用于粗核糖核酸(RNA)制剂或细胞裂解物,否则会堵塞标准的寡聚(dT)纤维素柱系统。本方法降低了降解风险,效率高,并且可以很容易地扩大或缩小规模。