what does isolating clones mean??

[kies89]

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for exampe, isolate cDNA clones,

what does that mean?

 

[Dotoday]

CDNA stands for complementary DNA. In eukaryotic cells DNA has introns and exons, introns are splices out anx exons are spliced in in eukaryotic cytoplasm. Since prokaryotes do not have a splicing machinery, you cannot express eukaryotic genes in prokaryotes. To go around this problem researchers make complementary DNA out of mature mRNA using reverse transcription PCR. mature mRNA has no introns. Now that we synthesized cDNA we can insert it in bacteria or yeast and make eukaryotic proteins. So, you isolate cDNA clones of cells that have your cDNA, you put that it in culture, now you can insert it in bacteria or yeast to make your proteins. Hope this helps.

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[biomolecmed]

When you infect bacterial cells with the same DNA, you let them grow in culture overnight to maximize the amount of cDNA that you have. Since all the cells are just repplicating, you essentially just making copies of those cDNA's. These are called clones, because they are the same sequence. When you purify the DNA from your bacterial culture, you also test it to make sure ththere are no unwanted mutations. This procedure is called isoating DNA cones. Hope this made it clear.