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Cloning involves insertion of genetic sequences in plasmids using techniques as restriction digest and PCR. However, traditional cloning techniques bring with it problems such as:
- Carry-over of salts and residual ethanol or other organic solvents that can inhibit downstream enzymatic reactions
- Multiple purification steps, each associated with significant product losses
Use of Nanosep centrifugal ultrafiltration devices in various steps of the cloning process offers the following benefits:
- Avoids retention of components such as ethanol or other organic solvents that may inhibit enzymatic activity
- High-yield rapid recovery of dilute DNA samples through reduced manipulations
We present a method for cloning PCR products or other DNA fragments that utilizes a single Nanosep centrifugal device and provides significant reductions in time, labor, and materials over classical methods.