IGEM:Groningen/Notebook/iGEM 2011/2011/05/26

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26-5-11


Protocols for transforming biobricks in cells on http://partsregistry.org/Help:Spring_2011_DNA_distribution

DNA Kit Plate Instructions
To use the DNA in the Distribution Kit you may follow these instructions:

1.With a pipette tip, punch a hole through the foil cover into the corresponding well of the Biobrickā„¢-standard part that you
want. Make sure you have properly oriented the plate. We recommend that you do not remove the foil cover, as it could lead to
cross contamination between the wells.
2.Pipette 10uL of dH2O (distilled water) into the well. Pipette up and down a few times and let sit for 5 minutes to make sure
the dried DNA is fully resuspended. We recommend that you do not use TE to resuspend the dried DNA.
3.Transform 1 or 2uL of the resuspended DNA into your desired competent cells, plate your transformation with the appropriate
antibiotic* and grow overnight. *
4.Pick a single colony and inoculate broth (again, with the correct antibiotic) and grow for 16 hours.
5.Use the resulting culture to miniprep the DNA AND make your own glycerol stock (for further instruction on making a glycerol
see this page). We recommend using the miniprepped DNA to run QC tests, such as restriction digests and sequencing.
* To know which antibiotics to use, look at the plasmid that the part is in. The naming scheme for plasmids is specifically
designed to indicate antibiotic resistance.

Note: There is not enough DNA in each well to perform anything but transformations

* Transformation:
put your cells+DNA 30min. on ice
incubate them for 2 min at 42 degrees
add 300 microliter LB medium and incubate them at 37 degrees
Plate 250 microliter and 50 microliter
incubate overnight at 37 degrees


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