How many rounds of PCR would be needed to amplify a DNA sequence by a factor of

Question

How many rounds of PCR would be needed to amplify a DNA sequence by a factor of approximately 64*10 12 ? Show a calculation. Hint: Remember, every round of PCR has a maximum amplification of 2-fold, and every ten rounds an amplification of approximately 1000-fold. So, to get an amplification of 4.000 one would do 2 + 10 = 12 rounds . This, of course, assumes high yields for each step.

6. You wish to insert a fragment of duplex DNA that is flanked on both ends by known restriction sites into a plasmid. What restriction site(s) would need to be present in the plasmid for this approach to work? If the plasmid were to be used to place the gene into a bacterium, outline the steps you would need to take and how you might detect if the transfer of genetic material was successful AND if the protein coded for were produced successfully. That is, how would you detect if the DNA fragment was inserted in the plasmid and transferred into the bacterium, and if the protein was expressed? How would you verify identity of the protein?

Explanation / Answer

If you want to know all the restriction enzyme for your DNA sequence then http://tools.neb.com/NEBcutter2/ can help you.

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