Lab Resources

PROTOCOLS

Frequently used laboratory protocols:

PREPARATION AND TRANSFECTION OF NICOTIANA MESOPHYLL PROTOPLASTS

TRANSIENT AGROINFILTRATION OF NICOTIANA LEAVES

TRANSFORMATION AND REGENERATION OF NICOTIANA

TYPE IIS MEDIATED PARALLEL DNA ASSEMBLY

We use a Type IIS parallel DNA assembly method (also known as Golden Gate Cloning). We have published a toolkit for plants as well as the PhytoBrick TypeIIS standard. Full details and a  laboratory protocol can be found in this methods chapter: Patron, NJ (2016). DNA assembly for plant biology. Current Protocols in Plant Biology 1:604-616

To help new users get started with Type IIS parallel assembly, we made a training video that explains the background behind easy and reliable assembly of multiple standard parts in a single reaction and how to make your own standard parts. Some of the laboratory protocols are slightly out of date now, but it’s a good place to start understanding the background. You can also download a help-sheet on how to make new parts.

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