Copenhagen/27 July 2011


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  • Ligate the the two sets of plasmids with the insert and make two controls to test if the digestion went well. One with only T4 buffer and one with T4 buffer and T4 ligase.
  • Colony PCR was done to 10 randomly selected colonies of B1 to screen for one with the right insert. Colony 1,9 and 10 showed vague signs of having this insert.

Overnight cultures of 1,9 and 10, which were made before knowing if they were any good, were purified and a sample from each was double digested with Ecor1 and PST1, whith the intention of determining whether or not the wanted insert was to be found.

  • Yesterday Casper did 3 ligations of A2. One was transformed after a 1 hour table ligation period. 3 Colonies grew from this transformation, these were transferred to another plate and dubbed A,B and C. From a colony PCR colony B showed signs of having the insert. An overnight colture was made of B.

The two other ligations which were overnight ligations were transformed.

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