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From 2011.igem.org

Wednesday

A few mistakes from last week, a fully booked PCR machine and a crowded freezer led to a morning of low spirits and around 12 O'clock it seemed there were nothing more that could be done. But after a lunch everything looked much brighter and positive thinking and troubleshooting kept us busy from then on - the A1 was ligated into the standard plasmid AND the expression vector. Around 2 O'clock another miracle happened as a missing BioBrick disappeared in the freezer and made sure that we had something to do for the rest of the day as well.

A1 The only one without any problems.

• Double Digestion with Xba1 and Pst1

• Ligation with the expression vector
  • Size of BioBrick J04500 = 220bp+3189=3409 bp
  • Size of BioBrick A1 = 1600bp
  • 3409/1600= ca. 2

• Transformation

Expressionvector'

• Double Digestion with Pst1 and Spe1