Which of the following is a primarily RNA based technique?
(A) Next generation sequencing (B) PCR (C) Sanger's technique (D) Western blotting
(B) PCR is Reverse Transcriptase PCR, where staing material is RNA. It is used to make cDNA, which is amplified. Thus RNA can be amplified by this method and quantification of mRNA can also be done (see fig). Here we use Tth polymerase enzyme which has both reverse transcriptase and polymerase activity WITH REGARD TO OTHER OPTIONS: Western blot is to detect protein. Next generation sequencing & Sanger's technique are for DNA sequencing. ADDITIONAL EDGE: Used to detect RNA expression Sample here is RNA. This RNA is conveed to cDNA by enzyme reverse transcriptase. Tth polymerase (Thermus thermophilus HB-8) has Polymerase as well as reverse transcriptase activity when mixed with manganese ions and can thus be used for the amplification of RNA to cDNA. This polymerase is thermostable but does not has proofreading activity. So it is usually combined with a proofreading enzyme.