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11. Why is the DpnI digestion necessary after performing the mutagenic PCR, and what sequence is...

11. Why is the DpnI digestion necessary after performing the mutagenic PCR, and what sequence is DpnI specific for?

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Answer #1

Step 1

Dpnl digestion is necessary after performing the mutagenic PCR because mutatents may be formed which must be digested by dpnl. Dpnl also digests methylated DNA which may be formed by mutagenic PCR. Dpnl digestion also destroys plasmid fragments.

Step 2

Dpnl is a type IIM restriction enzyme .

It's restriction site is

GmA|TC

CT| mAG

GmA| TC is also known as dam sequence.

Dpnl is highly specific for methylated DNA.

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