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Team:Grenoble/Biology/Protocols/BB
From 2012.igem.org
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| - | + | <h1>DNA Kit Plate Instructions</h1> | |
| + | </section> | ||
| + | <br/> | ||
| + | <section> | ||
To use the DNA in the Distribution Kit you may follow these instructions: | To use the DNA in the Distribution Kit you may follow these instructions: | ||
| - | + | <ul> | |
| - | + | <li>With a pipette tip, punch a hole through the foil cover into the corresponding | |
| - | : | + | well of the Biobrick™-standard part that you want. |
| - | + | <a href="http://partsregistry.org/Help:Spring_2011_DNA_distribution#DNA_Kit_Plate_Orientation"> Make sure you have properly oriented the plate</a>. | |
| - | + | We recommend that you do not remove the foil cover, as it could lead to cross contamination between the wells.</li> | |
| + | <li>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. | ||
| + | <li><a href="http://partsregistry.org/Help:Transformation_Protocol">Transform</a> 1 or 2uL of the resuspended DNA into your desired competent cells, | ||
| + | plate your transformation with the appropriate antibiotic* and grow overnight.</li> | ||
| + | <li>Pick a single colony and inoculate broth (again, with the correct antibiotic) and grow for 16 hours.</li> | ||
| + | <li>Use the resulting culture to <a href="http://openwetware.org/wiki/Miniprep/Kit-free_high-throughput_protocol"> miniprep</a> | ||
| + | the DNA AND make your own glycerol stock (for further instruction on making a glycerol see | ||
| + | <a href="http://openwetware.org/wiki/Endy:Making_a_long_term_stock_of_bacteria"> this page</a>). We recommend using the miniprepped DNA to run QC tests, such as restriction digests and sequencing.</li> | ||
''* To know which antibiotics to use, look at the plasmid that the part is in. The [http://partsregistry.org/wiki/index.php/Help:Plasmids/Nomenclature naming scheme] for plasmids is specifically designed to indicate antibiotic resistance.'' | ''* To know which antibiotics to use, look at the plasmid that the part is in. The [http://partsregistry.org/wiki/index.php/Help:Plasmids/Nomenclature naming scheme] for plasmids is specifically designed to indicate antibiotic resistance.'' | ||
''Note: There is not enough DNA in each well to perform anything but [[Help:Transformation_Protocol|transformations]]'' | ''Note: There is not enough DNA in each well to perform anything but [[Help:Transformation_Protocol|transformations]]'' | ||
| - | + | ||
</div> | </div> | ||
Revision as of 07:36, 24 August 2012
DNA Kit Plate Instructions
- With a pipette tip, punch a hole through the foil cover into the corresponding well of 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.
- 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.
- Transform 1 or 2uL of the resuspended DNA into your desired competent cells, plate your transformation with the appropriate antibiotic* and grow overnight.
- Pick a single colony and inoculate broth (again, with the correct antibiotic) and grow for 16 hours.
- 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 [http://partsregistry.org/wiki/index.php/Help:Plasmids/Nomenclature naming scheme] for plasmids is specifically designed to indicate antibiotic resistance.'' ''Note: There is not enough DNA in each well to perform anything but [[Help:Transformation_Protocol|transformations]]''
