Question

In conventional molecular biology experiments, DNA isolation is only the first step. What are the next possible steps in a typical research setting? Cite examples if possible

Answer 1

After DNA isolation,the DNA is digested with some specific of the restriction enzymes and then it is run in agarose gel electrophoresis then the specific fragments are hybridized by using specific probes (flourescent or radiolabelled) which are bind to DNA specific fragments.These fragments obtained from southern blotting can be used in molecular cloning experiments where these specific DNA fragments are introduced into vector that is plasmid to produce recombinant clones.

Another experiment is RFLP experiment which is a restriction fragment length polymorphism. For example to know the SNP for causing asthma disease then first DNA should be isolated and then the next step is to set up a PCR reaction by designing specific primers,after that prepare reaction mixture that is ( dNTP's,taqbuffer,taq polymerase,distilled water,and primers).Add the template that is DNA which is isolated and the run pcr under appropriate annealing conditions.Then load into gel the products of pcr are then restricted digested using restriction enzymes which are specific.Then run the gel electrophoresis where the bands are fragmented based upon the specific polymorphism.