IGEM:Groningen/Notebook/iGEM 2011/2011/05/17: Difference between revisions
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== | ==17-5-11== | ||
* | <br> Biobricks arrived today! | ||
<br> Protocols for transforming biobricks in cells on http://partsregistry.org/Help:Spring_2011_DNA_distribution | |||
<br> | |||
<br>DNA Kit Plate Instructions | |||
<br>To use the DNA in the Distribution Kit you may follow these instructions: | |||
<br> | |||
<br>1.With a pipette tip, punch a hole through the foil cover into the corresponding well of the Biobrick™-standard part that you <br>want. Make sure you have properly oriented the plate. We recommend that you do not remove the foil cover, as it could lead to <br>cross contamination between the wells. | |||
<br>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 <br>the dried DNA is fully resuspended. We recommend that you do not use TE to resuspend the dried DNA. | |||
<br>3.Transform 1 or 2uL of the resuspended DNA into your desired competent cells, plate your transformation with the appropriate <br>antibiotic* and grow overnight. * | |||
<br>4.Pick a single colony and inoculate broth (again, with the correct antibiotic) and grow for 16 hours. | |||
<br>5.Use the resulting culture to miniprep the DNA AND make your own glycerol stock (for further instruction on making a glycerol <br>see this page). We recommend using the miniprepped DNA to run QC tests, such as restriction digests and sequencing. | |||
<br>* To know which antibiotics to use, look at the plasmid that the part is in. The naming scheme for plasmids is specifically <br>designed to indicate antibiotic resistance. | |||
<br> | |||
<br>Note: There is not enough DNA in each well to perform anything but transformations | |||
<br> | |||
<br> * Transformation: | |||
<br> put your cells+DNA 30min. on ice | |||
<br> incubate them for 2 min at 42 degrees | |||
<br> add 300 microliter LB medium and incubate them at 37 degrees | |||
<br> Plate 250 microliter and 50 microliter | |||
<br> incubate overnight at 37 degrees | |||
Revision as of 02:02, 1 August 2011
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17-5-11
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