Subcloning Tips - very important heating step
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Subcloning Tips - very important heating step |
| Source: Schimmelpenninck |
| Abstract: Put your insert and vector in an Eppendorff tube and then heat it for 5 minutes, then add ligation buffer and ligase |
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After isolating your digested fragments from the gel put digested vector and digested insert TOGETHER in an Eppendorff tube, then put this for about 5 minutes at 65 degrees celsius, then cool it and spin it down, then add your ligation buffer and ligase. The 65 degrees celsius step is VERY important.
Short explanation: |
