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Team:TU Darmstadt/Notebook/Labjournal/K1497031
From 2014.igem.org
Scaffold with His-tag
The standard PCR for the generation of the insert yielded a clear band of the correct length (see figure 1). The amplicon should have a size of 943 bp, which corresponds with the height of the received band. After ligation and transformation in E. coli Top10 cells, existing colonies were examined via colony PCR (see figure 2). The desired insert was present in two of three tested clones. After isolation of the plasmid DNA, the successful cloning procedure was verified by sequencing.
Figure1: Colony PCR of the scaffold protein with His-tag (SH) cloned into pSB1C3. The 2-log ladder was applied onto the left side. The PCR product should have a length of 943 bp.
Figure 2: Standard PCR of the scaffold protein with His-tag (SH) for molecular cloning into the vector pSB1C3. The 2-log ladder (M) was applied onto the right side. The PCR product has a length of 943 bp.
