Part:BBa_K2560001:Experience

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Experiences of working with the Marburg Collection in E. coli

We, the iGEM Team Heidelberg 2020, really liked working with the Marburg Collection in Golden Gate Cloning and want to thank the iGEM team Marburg 2018 for creating such a great library of biobricks. We successfully showed that the Marburg Collection can be used in E. coli as well. For our work, we adapted the protocols of the iGEM Marburg Team 2018 (especially in the colony PCR) and developed a new protocol for another ligase, the NEB Hi-T4 Ligase.

Futur iGEM teams we want to give the following tips:

A lot of thoughts should be put into the primer design.

In Level 0 cloning, only 7.5 ng of the entry vector should be used.

The resistance gene should always be added in a molar concentration 1/10-fold compared to the other components in Level 1 and 2 cloning.

Cells do not have to be lysed for colony PCRs.

Loooooots of colonies should be picked (Yey picking parties!).

For more information about this, please check out our wiki.

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