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Usage and extraction

Usage

The Distribution Kit Plates contain dried DNA of hundreds of parts that are available in the Registry up to 2009. While there is not enough DNA for assembly, you will be able to transform the DNA into cells and then make your own glycerol stocks of any part that you wish. [edit] Linearized Plasmid Backbone Instructions

In addition to the DNA plates the Distribution Kit also contains a set of linearized plasmid backbones:

• pSB1C3
• pSB1T3
• pSB1A3

These plasmid backbones have been prepared via PCR and purified. All you need to do is cut with EcoRI, PstI and DpnI to leave two ends ready to be ligated to a Biobrick™ part. Use these backbones to send your parts to the Registry when shipping your parts before the Jamboree.

• NOTE: The linearized plasmid backbones need to be cut by EcoRI and PstI restriction enzymes prior to use.

You can find instructions on how to use AND make your own linearized plasmid backbones on the protocol page. [edit] 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 to the Biobrick™-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. Add 10uL of diH2O (deionized water)

3. Pipette 1 or 2uL of the resuspended DNA transform into your desired competent cells, plate bacteria with the appropriate antibiotic* and grow overnight.

4. Pick a single colony and inoculate broth (again, with the correct antibiotic) and grow for 18 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.

--Rchamberlin 19:51, 28 July 2010 (UTC)