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Your methods so far have given some success! You now want to look at many different tissues in several individuals, some of whom have the mutation, some who are controls. How can you efficiently generate and amplify cDNA from each individual and tissue that can be used for directional cloning? Remember, you want to do many samples from many tissues, so starting with total cellular RNA fractions containing mRNA is where you need to put your effort and resources!
In the eukaryote cell nucleus, RNA can be found dispersed in the nuclear fluid, along with DNA, and as the major constituent of the nucleolus. In cytosol (in eukaryotes or in bacte
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