Team:TU Darmstadt/Notebook/Labjournal/K1497005

From 2014.igem.org

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K1497005 - Composite part of CHS and CHI with strong RBS (B0034-CHS-B0034-CHI)

For combing two of our Biobricks, we digested B0034-CHS-pSB1C3 with EcoRI + SpeI and B0034-CHI-pSB1C3 with EcoRI + XbaI. Furthermore we dephosporylated the digestion of B0034-CHI-pSB1C3. We ligated both digestions with T4 ligase and transformed the product into Top10 E.coli via heat shock. We spread out on CMP-LB agar plates and incubated the plates overnight at 37°C. We analyzed obtained colonies by colony PCR and agarose gel electrophoresis. Positive colonies were inoculated in 5 ml LB with CMP and plasmids were isolated after incubating for 16 h at 37°C gaining the BioBrick BBa_K1497005.