NORTHERN BLOTTING (BIOTECHNOLOGY)
This blotting technique is mainly used for identification of RNA molecules.
The
steps involved in this method are almost similar to that of Southern blotting.
Northern blotting developed in the year 1977 by James Alwine, David Kemp,
and George Stark at Stanford University, USA.
DNA blotting is named after its
inventor
Southern, this technique has got the name as Northern owing to the
similarity in blotting and easy to memorize.
Northern blotting, include: isolation of RNA, electrophoresis,
followed by
membrane transfer, hybridization and performing autoradiography.
RNA
molecules highly sensitive towards RNase enzyme that is widely available in
the environment even on the tips of our fingers.
Northern blotting to prevent contamination and loss of RNA
sample.
Moreover, the isolated RNA would be in the secondary confirmation,
to destabilize this secondary structure formaldehyde is used.
In the early days,
researchers were using diazo benzyloxymethyl (DBM) paper for transferring
the RNA.
However, these days researchers using modified nylon membrane
instead of DBM papers.
After transferring the RNA molecules over to nylon membrane, they are
subjected to hybridization using DNA probes. Later autoradiogram was
developed by using autoradiography method..
Northern blot in is
extremely useful for studies like gene expression in tissues and specific cell
lines. One of the major limitations of this technique is, “it measures the
accumulation of RNA transcripts rather than individual RNA”
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