Team:USTC CHINA/Notebook/Protocols/Overlap PCR
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<h1>Overlap PCR</h1> | <h1>Overlap PCR</h1> | ||
- | <p>The application of overlap PCR is widespread, and the mechanism is actually simple: suppose you are going to link two gene or a promoter and a gene, the primary method is cutting out by enzyme, but frequently we fail to find suitable restriction enzyme sites, or the enzyme is too special or expensive, in either situation purchasing this enzyme can turn out to be a waste. Overlap PCR affords a better alternative. Here is an illuminating example: suppose we have two genes, namely A and B, whose sequences are 5-atgcatgctagctagaacgctacgctgactaccccctgatc-3 and 5-atgctagtagctagccccccccaggggataattttttaaaacg-3 separately. At the beginning we should design primer, and the sequences of primer is assumed as follows:</br> | + | <p>The application of overlap PCR is widespread, and the mechanism is actually simple: suppose you are going to link two gene or a promoter and a gene, the primary method is cutting out by enzyme, but frequently we fail to find suitable restriction enzyme sites, or the enzyme is too special or expensive, in either situation purchasing this enzyme can turn out to be a waste. Overlap PCR affords a better alternative. Here is an illuminating example: suppose we have two genes, namely A and B, whose sequences are</br> 5-atgcatgctagctagaacgctacgctgactaccccctgatc-3 and 5-atgctagtagctagccccccccaggggataattttttaaaacg-3 separately. </br>At the beginning we should design primer, and the sequences of primer is assumed as follows:</br> |
A1:5-ATGCATGCTAGCTAGAACGCT-3</br> | A1:5-ATGCATGCTAGCTAGAACGCT-3</br> | ||
A2:5-ggggggctagctactagcatgatcagggggtagtcagcgt-3</br> | A2:5-ggggggctagctactagcatgatcagggggtagtcagcgt-3</br> |
Revision as of 17:34, 25 September 2013