The DIG System - a High Sensitive Substitute of Radioactivity in Northern Blot Analysis

摘要

The Northern blot technique is frequently employed for the characterization of gene expression. RNAs and for gene function analysis. Using radioactively-labeled probes is still a common technique in Northern blot analysis. The main disadvantages of this technique are the possible health hazard, inconvenience during handling and the short half-life of the probe. Here we demonstrate that the digoxygenin (DIG) system is an easy and very sensitive alternative in Northern blot analysis. DIG-labeled probes are stable for at least one year and guarantee short detection times. By analyzing the transcription pattern of the mRNAof the human DEAD box protein p68 RNA helicase [1] in different tissue types it is shown that DIG labeling is comparable to radioactive labeling. Furthermore an optimized stripping protocol opens up new possibilities for stripping and reprobing of Northern blots.