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one tube cloning method

One-Tube PCR Cloning Method

Chun-Ming Liu

This method has 3 reactions (restriction, polishing and ligation) carried out in one tube using ligation buffer. Believe or not, it works very well for me and a lot of my friends.
  1. Purify PCR product using Spin-Column S-300 from Pharmacia.
  2. Ligation mixture (you may be able to reduce the cost by reducing the volume):
  3. Transform directly to DH5a ultra-competent cells using my favorite 5 minutes method.

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