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Northern Analysis: The Basics

probe. This process can be time consuming and problematic, since harsh treatment is required to strip conventional probes from blots. Ambion has addressed this limitation with the advent of the Strip-EZ StripAble RNA, DNA and PCR Probe Synthesis and Removal Kits. These kits produce high specific activity RNA and DNA probes that can be quickly and easily stripped from blots using mild conditions.


Steps Involved in Northern Analysis

Although established Northern blotting procedures are up and working in most molecular biology laboratories, Ambion has found ways to considerably improve on standard protocols, resulting in greatly increased Northern sensitivity. We have developed RNase-free reagents optimized for each step of the procedure (Figure 1) to provide complete, high-sensitivity Northern blotting systems.

The steps involved in Northern analysis include:

  • RNA isolation (total or poly(A) RNA)
  • Probe generation
  • Denaturing agarose gel electrophoresis
  • Transfer to solid support and immobilization
  • Prehybridization and hybridization with probe
  • Washing
  • Detection
  • Stripping and reprobing (optional)

Figure 1. Northern Analysis with Ambion Kits.


RNA Isolation

Obtaining high quality, intact RNA is a critical step in performing Northern ana
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