Megaprimer based PCR Flashcards Preview

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Flashcards in Megaprimer based PCR Deck (3)
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1

What is megaprimer PCR?

Where you use a larger primer than normal to introduce novel sequences to the template DNA

2

What novel sequences can be added?

Flanking regions (RE sites)
Upstream nucleotides
Modified terminal codons
Leader, linker, and tag sequences
Overlap sequences

3

Describe the primers used?

Very long- up to 300bp
Non-complementary 5' sequences
Contain the novel sequences to be added in the overhang part of the primer
The 3' complementary side has its Tm calculated but no the rest of the sequence