北方墨點法

北方墨點法（英語：Northern blot），又称 RNA 印迹法，是分子生物学、生物化学中常用的实验方法，通過探測樣品中的RNA（或隔离的mRNA）来研究基因表达。[1][2]

通过北方墨點法来检测RNA的流程图

利用探針偵測由凝膠電泳分離出來的RNA片段，尋找含有特定序列的RNA片段。與南方墨點法相似，RNA樣品經過凝膠電泳後會按照大小分離，然後將樣品從凝膠轉印到膜上，並用與目標序列互補的標記的探針來探測。實驗結果可以根據所用探針的不同，以多種方式來觀察，但大多數都顯示的是樣品中被探測的RNA條帶的相對位置，也就是分子大小；而條帶的強度則與樣品中目標RNA的含量相關。這一方法可以測量目標RNA在不同樣品中的情況，因此已經被普遍用於研究特定基因在生物體中表現的時刻和表現量，也是這類研究中最基本的手段。

發明者是史丹佛大學的James Alwine和喬治·斯塔克（George Stark）在1977年發明，其名字是源於第一種墨點技術，南方墨點法（命名者為Edwin Southern）[1]，兩者很相似，與南方墨點法主要的不同是北方墨點法探測的是RNA分子，而不是DNA分子。[3]

原理

與南方墨點法相似，膠體可以選用瓊脂凝膠或是聚丙烯醯胺凝膠，被檢測物是RNA，「探針」(probe)是與目標序列互補的RNA、DNA序列或是寡核苷酸，但是探針至少要有25個鹼基對與目標序列互補。

Capillary blotting system setup for the transfer of RNA from an electrophoresis gel to a blotting membrane.

膠體

RNA樣品通常以含甲醛的瓊脂凝膠電泳來分離，甲醛作為RNA變性劑，將RNA的二級結構變性成一級結構。凝膠內用溴化乙錠(EtBr)染色，可在UV光下觀察到RNA樣品的品質、大小。

實驗步驟

實驗大至分為鑄膠、跑膠、轉漬、北方雜合反應等幾個部分。

參見

参考文献

  1. ^ 1.0 1.1 Alberts, B., Johnson, A., Lewis, J. Raff, M., Roberts, K., Walter, P. 2008. Molecular Biology of the Cell, 5th ed. Garland Science, Taylor & Francis Group, NY, pp 538–539.
  2. ^ Kevil, C. G., Walsh, L., Laroux, F. S., Kalogeris, T., Grisham, M. B., Alexander, J. S. (1997) An Improved, Rapid Northern Protocol. Biochem. and Biophys. Research Comm. 238:277–279.
  3. ^ Bor, Y.C.; Swartz, J.; Li, Y.; Coyle, J.; Rekosh, D.; Hammarskjold, Marie-Louise. Northern Blot analysis of mRNA from mammalian polyribosomes. Nature Protocols. 2006. doi:10.1038/nprot.2006.216. 

外部链接

關於北方墨點法
的圖書館資源
维基共享资源上的相关多媒体资源：北方墨點法

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