Requirements for DNA Submission

Our service of mouse creation via DNA microinjection starts with the importation of the DNA sample. To expedite the delivery of your project, make sure to provide all the items listed below.

Submit these items for plasmid DNA microinjection:

• At least 50µg of purified plasmid DNA of sufficient purity for restriction digest in a labeled tube on cold packs
• A map of the plasmid with relevant restriction enzyme sites denoted
• Clear instructions on which enzyme(s) to be used for releasing transgene from vector or linearizing plasmid
• Size of plasmid and expected fragment sizes(s) post-digestion
• An image of the digested plasmid run on an agarose gel with a DNA ladder (molecular weights indicated)

Submit these items for BAC DNA microinjection:

• BAC DNA or a glycerol stock in a labeled tube
• At least 10µg of BAC DNA on cold packs or a glycerol stock on dry ice
• BAC growth conditions:
• Growth temperature
• Antibiotic resistance
• Medium composition

Important information regarding DNA preparation for microinjection

• Linearized transgene DNA or supercoiled BAC DNA will be microinjected into fertilized embryos for the production of transgenic mice.
• We verify the concentration of the DNA before and after processing. If insufficient quantities of DNA are present, we will report this to you and stop any further work until the issue is resolved.
• Transgenes for microinjection should be free of plasmid vector sequences. The design of your transgene should include restriction enzyme sites that result in fragments that can easily be separated from the prokaryotic backbone by agarose gel electrophoresis.
• We make up to two attempts to digest plasmid DNA. If the vector does not cut with the suggested enzymes under standard conditions, or does not result in the expected fragment size(s), we will report these results to you and stop any further work until the issue is resolved.

The Jackson Laboratory cannot accept transgenic service projects that will require laboratory or animal room procedures at the BSL2/ABSL2 level or higher. For example, we will not make transgenic mice from lentiviral or adenoviral vectors, either by infection or DNA microinjection, nor can we house mice that may express any transgene or gene product that requires the mice to be housed or handled under BSL2 or ABSL2 conditions.