A researcher chooses a method that is simple and fast for TMV resistance detection. Which is best?
Random amplified polymorphic DNA offers rapid, inexpensive screening without requirement for sequence information, radioactive probes or sophisticated instrumentation. Using a single short random primer in a standard PCR, numerous loci are amplified simultaneously, producing easily scorable fingerprints on agarose gels within hours. For preliminary disease resistance screening such as Tobacco mosaic virus, quick identification of linked fragments is advantageous. RFLP and AFLP involve laborious digestion, blotting and selective amplification steps, while EST-SSR development requires transcriptome sequencing and primer validation, increasing cost and time.
Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.