Ribotyping, typify bacteria by RNA analyses | see ARDRA also
Channel: Dr. Sanjeev Pandey
Date: 2023-10-01

Ribotyping – Ribotyping is a molecular technique used to identify unknown bacterium. It is basically the RFLP (Restriction fragment length polymorphism) of rRNA genes. The banding pattern of the unknown bacterium is compared with that of standard bacteria that are used as reference. The process starts with the isolation of the whole genome first, followed by restriction digestion with a restriction endonuclease. The digested DNA is then run on an agarose gel. Same procedure is followed for the standard bacteria and their DNA is loaded in the consecutive lanes in the gel. This is followed by southern blotting and hybridization using a probe of rDNA region. The rDNA probe (radiolabeled oligonucleotide of the rDNA element) finds its complementary target DNA in the membrane and hybridizes there with. After that the membrane is washed in SSC (Sodium citrate- Sodium chloride) buffer at appropriate stringency  to remove the background (non specific binding of the probe). Following this the membrane is exposed to X-ray film, a process called autoradiography.  The autoradiogram thus produced reveals a specific banding pattern characteristic of a species. For identification, the banding pattern of query DNA is matched with the standard DNA of reference bacteria in the autoradiogram.   
Also see the blog: https://spbio-asnwb.blogspot.com/2023/12/blog-post.html
For details See book - Advanced Botany VolII By Sanjeev Pandey
https://getmybooks.com/advanced-botany-volume-2/p/b949c15965