Team:TU Darmstadt/Notebook/sec1/K1602036
K1602036 - T7-Acetyl Esterase-SH3-ligand (T7-aes-SH3Lig)
K1602034 was digested with XbaI and PstI, ligated into the backbone T7-pSB1A2 and transformed into E. coli Top 10 cells. Afterwards, colonies were screened by colony PCR (oligonucleotides VF2 and VR) (Figure 1). The plasmids of positive colonies were extracted and digested with EcoRI and PstI. The obtained fragment was cloned into pSB1C3 and transformed into competent E. coli Top10 cells. Colonies were screened by colony PCR (oligonucleotides VF2 and VR) to verify positive colonies.
Figure 1 Colony PCR of T7-aes-SH3Lig (1-4). The size of the amplified product was around 1.4 kbp. DNA marker: 2-Log DNA Ladder. (NEB).