Service Request Procedure

  1. Please contact Zhanyuan Zhang via e-mail (zhangzh@missouri.edu), by phone (573-882-6922), or meet in our transformation lab to discuss your service request.

  2. After the initial contact, we will send you an invoice for payment. After receiving the payment, we will start transformation experiments. If you are located outside the state of Missouri, please contact USDA-APHIS (http://www.aphis.usda.gov/) as soon as possible, to request permission for cross-state movement of transgenic materials.

  3. You will need to subclone your gene expression cassette into our binary vector pZY101, pZY101.2, pFGC5941(-), pFGC5941 or other vectors published by PTCF. Off-campus users are required to sign a Material Transfer Agreement sheet to use any of these vectors. We will mobilize the vector into Agrobacterium cells and confirm its integrity within the cells at a charge of $130/vector confirmation. Please provide the restriction map of your construct for our confirmation work.

Service Note

Below is a list of what we would like to communicate with our users so the requirements and expectations will be clear before the service experiments start.

  1. To minimize costs for both users and our facility, an average number of events per construct will be generated for users. For example: if users require 10 events per construct we will generate an average of 10 events per construct, instead of at least 10 events per construct. However, the probability of the number of events less than users require will be minimal.

  2. Users are encouraged to request empty vector control events. This will not only benefit the user’s publication requirement but also allow evaluating if user’s genes of interest have any negative impact on plant transformation in our side-by-side service experiments.

  3. The facility's responsibility is to warrant the presence of genes of user's interest but not the expression level. This expectation is based on the complexity of gene expression and the user's choice of gene expression systems.

  4. Standard time for service deliverables: The normal service deliverables are: the first young seedling of transgenic soybean event will become ready for sampling at about five months, and dry seeds of first transgenic event will be ready in about eight to nine months after the first service experiment starts. Subsequent delivery will take place in additional months because each transformation experiment takes at least six months from start to completion. The standard delivery time of maize service deliverables takes even longer because of the longer tissue culture period required for maize somatic embryogenesis. The standard delivery time of switchgrass and sorghum events ready for PCR or Southern blot analysis are around 6 months.
  5. When users confirm the vector integrity within the Agrobacterium, PCR-based confirmation will no longer be considered as a valid proof, and users are highly recommended to show digestion of the final transformation vector rescued from bacterial cells carrying the gene of interest. This new requirement will benefit users as well as our operation because of more thorough examination of those steps before the transformation experiments start.

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