Northern Blot Analysis of microRNAs and Other Small RNAs in Plants
The study of regulatory small RNAs, such as siRNAs and microRNAs in plants, has necessitated methods tailored to their unique features. Their analysis demands the use of sensitive and quantitative methods for their detection. The use of Northern blot hybridization offers an attractive alternative to address qualitative as well as quantitative features. We highlight the advantages and shortcomings of this method and offer a detailed description of the techniques that best work in our hands, considering their use for the study of several small RNAs in multiple samples. We enumerate relevant details as well as cautionary comments in cases where we have detected potential difficulties.
Key wordsPlant sRNAs Plant miRNAs Northern blot
CDR received a PhD fellowship from CONACyT. We are grateful to Daniel Blanco-Mello for kindly providing the Northern blot image shown in Fig. 1 (DBM and JLR, unpublished results). This work was partially supported by grants from UNAM (PAPIIT IN-205015, IN-202918) and from CONACyT (CB2010-151571) to JLR.