Transformation: 4500 SEK
Includes transformation and propagation of about 10-14 individual lines (“1st mother Jar”)

Routine cut:
Internal: 50 SEK / Jar
External: 80 SEK / Jar

Amplification:
Internal: 50 SEK / Jar
External: 80 SEK / Jar

Propagation of T89, Elite 864012 or Tobacco:
Internal: 60 SEK / plant
External: 80 SEK / plant

Routine cut after 3 years:
Internal: 120 SEK / Jar
External: 150 SEK / Jar

New transformation if the transformation fails:
During summer (midsummer to mid-august): 1500 SEK
Rest of the year: a failed transformation is redone once for free

Forgotten transformation: 400 SEK
(We still have to prepare and we lose at least one hour work)

Cut in 50 ml falcon tube for transport:
Internal: 40 SEK

Samples:
35 SEK / Sample (tube)

Transformation

To request a transformation you first need to fill in the form found under “request transformation” which you find here to the right. Please follow the guidelines stated there.

If you want to make a CRISPR CAS9 transformation we will treat that as 2 transformations which means we will have double amount of explants and you will also receive double amount of lines. Please try to put the name CRISPR in the name of the construct to avoid any misunderstanding. Regardless if you want a regular or CRISPR transformation. You need to provide the Agrobacterium containing the construct of interest as a pellet from a 50 ml liquid growing culture with an OD between 0.3-1.0 (max 1.5). The pellet must be provided in the morning of the given transformation day (preferably before 9.30 am).

The facility will then do:

• the transformation (regular 3 days), (CRISPR 3 days)
• selection on plates (Regular 2-3month),(CRISPR 2-6 month)
• growing the callus (Regular 1-1,5month), (CRISPR 1-2 month)
• growing the shoots (Regular 1.5-2month) (CRISPR 1,5-2month)
• selection of the shoots to get the individual lines (regular 1-2 month) (CRISPR 2-3 month)

If you forget your transformation appointment, we will charge for 1 hr work (400 SEK) (this is the time for us to prepare for the transformation in vain and putting everything back)

Selections available:

• Kanamycin
• Hygromycin

We can do transformations all year long but during the summer (midsummer to mid august) we do not redo the transformations for free if they get infected.

Amplifications

Download the pdf here to the right and fill in the request of amplification, print it out and put it in the basket on the 4th floor. State both how many plants you want and how many you want in each jar; the default is 3 plants per jar. Also remember to fill in if you would like wild type plants, and in that case, how many. Usually when starting an amplification, we only have 3 shoots to start with, so depending on how many replicates you want it might need several rounds of propagation. One propagation round takes normally between 4-5 weeks to grow. So please plan. We will try to inform you when we start the amplification and when we expect it to be ready.

Taking samples for testing

We will inform you when your transgenic lines are ready to be tested. There are two possibilities:

1. We can take the samples (stem or leave) for you as soon as the first individual lines are ready and freeze the samples in liquid nitrogen. The price for that is 35 SEK per sample
2. The other possibility is to wait a few months more until the next routine cut and then you can get the old jars. The disadvantage of that is that we have less material to start with if you order an amplification of your selected lines.

For CRISPR transformation we take the samples by default in an earlier step so if you do not want samples you have to tell us in advance.

Procedure for transformation request

• Fill in the request below. Choose the construct name carefully, so it will be possible to identify the construct in the database in the future, both for yourself and for others.
• Print a paper copy of the confirmation of the request (use the browsers print function)
• Place the paper copy in the plastic basket, marked request forms, on floor 4 outside the greenhouse.
• Your transformation will be given a database number by Susanne Larsson.
• You will be contacted by a person in the transformation facility team, and you can together agree on a date when the transformation will be done. We will decide within the team who will be the responsible person.

Use your normal UMU-ID or SLU-ID login to the Request Form

There is no exception from this procedure. The transformations will be done in order.
Please, do not go and talk to any of us and ask to go before in the queue. That is not fair to your fellow colleagues and not fair to us.

Reverse genetic approaches are the only realistic alternative for the analysis of gene functions in trees because of their long generation time and obligatory out-crossing of most trees. UPSC scientists were among the first in the world to be able to create transgenic trees.

Poplar in vitro room 

All initial work was done with a fast-growing and relatively easily transformable hybrid aspen (Populus tremula x tremuloides) clone originating from the Czech Republic called “T89”. This is still the main tree species used in the transformation facility, but the numbers of different tree species that are routinely transformed have now grown to include elite varieties of hybrid aspen, Populus trichocarpa, Populus alba, Populus tremula and also Betula pendula (silver birch).

The method used is Agrobacterium-mediated plant transformation of stem segments, petioles or leaf explants and the facility is today growing more than 15 000 transgenic trees, representing more than 5000 different lines, probably making this the largest tree transformation facility in the world. The transformation facility currently employs five fulltime persons and is servicing all the research groups at UPSC and collaborators at both Umeå University and the Swedish University of Agricultural Sciences. Any group requiring transformed trees simply needs to provide the Agrobacterium strain carrying the genetic construct they want to analyze. After 8-10 months the trees are ready to be transferred to the greenhouse for analysis.

Different stages of poplar transformation: from explants to small trees on soil.

The persons working at the facility:

Contact:

For practical questions e.g. regarding the transformation procedure, please contact:
Veronica Bourquin at This email address is being protected from spambots. You need JavaScript enabled to view it.

For other questions, please contact:
Ove Nilsson at This email address is being protected from spambots. You need JavaScript enabled to view it.