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From 2010.igem.org

Andreas

Preparation of Top10 chemically competent cells

None of the clones streaked onto the Amp 100 plate (11/9) grew, indicating no AmpR contamination in cells. One of the ON cultures were therefore selected for preparation of competent cells.

Procedures according to protocol. Growth conditions changed to 25 °C, 220 rpm (OD₆₀₀ reached after ≈6 h).

A 100 μl aliquot was divided and spread onto Amp 100, Cm 25 and Km 50 plates to verify that they were not contaminated.

Cloning of N-CPPs into pSB1C3

Sequencing results from 8/9 returned.

• pSB1C3.nCPP 3 (failed)
• pSB1C3.nCPP 4 (fasta)
• pSB1C3.nCPP 6 (fasta)
• pSB1C3.nCPP 7 (fasta)
• pSB1C3.nCPP 8 (fasta)

Blastn alignments against N-Tra10, N-TAT and N-LMWP indicated successful cloning of N-Tra10 (clone 7) and N-TAT (clone 4).