What to do with a new clone
Yay! You made a new clone. Good work. Here are the several important steps you need to take next.
Register the Clone in the Registry of Standard Biological Parts
- Go to the Registryand choose "Add a part"
- Decide whether the new part is a Basic Part (discrete function units that cannot be subdivided, like promoter, RBS, coding sequence), Composite Part (functional unit composed of basic parts, like gene expression cassette, inverter), or Construction Intermediate (no function, byproduct of construction).
- Choose the number for the next available part
- Be sure to include short and long descriptions of the part
- Once data is available on the part, go back and include it as "Experience"
Enter the Clone into GCAT-alog
- Enter the information for each frozen tube into our online freezer stock database called GCAT-alog.
- Be sure to include all the requested information
Make a Glycerol Stock
In order to preserve your new clone for use by others for years to come, you need to make a glycerol stock (see: Glycerolstocks_How_to_Make_Glycerol_Stocks_of_Bacteria ). Do this right away. Bacteria on plates stored at 4 degrees survive for about 1 month. Bacteria in liquid cultures survive about 2 days on the bench and about 1 week at 4 degrees. Plasmids degrade rapidly in cells left in liquid culture.
- Grow a fresh overnight culture of the clone in the appropriate media with antibiotic
- Get a cryotube, cap, and colored cap insert. Write the part number on the color cap insert. Write the part number, the plasmid type (pSB1A2, pSB4T3, etc.), and a short description on the tube.
- Transfer 300 ul of glycerol into the labeled cryotubes.
- Add 700 µL of the overnight bacterial culture. Put the cap on and shake VERY vigorously, or vortex vigorously.
- Put the tube in the numbered freezer box location that was determined by GCAT-alog.
- When you want to retrieve some cells from the glycerol stocks, do not let them thaw. Use a sterile toothpick and scrape off some of the ice and put this onto your plate or into a liquid culture with the appropriate antibiotic.
Put some of the Plasmid in the Current iGEM Box
- Label a 1.5 ml tube with the part number, plasmid, and a short description of the part
- Place 10 ul of a miniprep of the plasmid into the tube. The concentration should be at least 40 ng/ul
- Place the tube in the freezer box labeled with the current year and "iGEM Registry Clones"