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<h4><strong><I>D.The SOE(Gene splicing by overlap extension) PCR</I></strong> </h4> | <h4><strong><I>D.The SOE(Gene splicing by overlap extension) PCR</I></strong> </h4> | ||
− | <p>We use SOE to | + | <p>We use SOE to ligate different genes, which are difficult to cut or ligate. We achieve it by overlap extension during PCR. |
First, we need to design 4 primers. Primer 1 & 4 are normal ones. The left part of primer 2 is normal primer, while the right part is primer for gene B. Primer 3 is just like primer 2. The right part of primer 3 is normal, while the left part is primer for gene A. Some base of primer 2 and primer 3 can reach complementary base pairing. | First, we need to design 4 primers. Primer 1 & 4 are normal ones. The left part of primer 2 is normal primer, while the right part is primer for gene B. Primer 3 is just like primer 2. The right part of primer 3 is normal, while the left part is primer for gene A. Some base of primer 2 and primer 3 can reach complementary base pairing. | ||
Second, we amplify gene A by primer 1 & 2 and amplify gene B by primer 3 & 4 respectively. After that we can get extended gene A and gene B. | Second, we amplify gene A by primer 1 & 2 and amplify gene B by primer 3 & 4 respectively. After that we can get extended gene A and gene B. |
Revision as of 18:48, 1 November 2017