Standardization of Northern blot non-radioactive protocol used in the detection of small RNAs in Vero line cells

Abstract

The interest in detection, identification and functional characterization of small non-coding RNAs (snRNAs), has generated the need to optimize the methodologies commonly used in its detection in specificity and sensitivity, The Quantitative reverse transcription PCR (RT-qPCR) and Northern blot. Even though the low sensitivity of Northern blot, this method continues to be commonly used in the sRNAs, because its capacity to detect the sRNA and its precursor, which is the reason why Northern blot is used as complementary method in this sort of Research. This work describes the implementation of an innovative non-radioactive Northern blot protocol, with modifications that improving the sensibility and specificity, with the discussion of the theoretical foundations of such modifications.

Keywords

• biología molecular
• técnicas moleculares
• investigación
• 1-ethyl-3-(-3-dimethylaminopropyl) carboniimide)
• Digoxigenine
• Northern blot non-radioactive
• LNA probe
• sRNAs