Team:Todai-Tokyo/Protocols/Infusion

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Infusion Description

Infusion is a method to combine DNA fragments using a combination of PCR and homologous recombination. It can be used as an alternative to ligation with the advantage that it does not require restriction enzyme digests, although specific primers are required for the reaction.

[http://bioinfo.clontech.com/infusion/ Clonetech page on In-Fusion]

Infusion Protocol

  1. Mix the purified PCR insert(50-200ng) and linearized vector(100-400ng)
  2. Add 1µl of 5x In-Fusion Reaction Buffer and MilliQ up tp 4.5µl
  3. Add 0.5µl of In-Fusion Enzyme and mix well

Incubate at 17ºC for 30 min.

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