Services:
Agrobacterium mediated maize inbred line B104 genetic transformation
Protocol: optimized based on Kang et al. (2022)
Target tissue: immature embryos
Transformable genotype: B104
Selection: the NPTII gene / G418
Product available: plantlets
DNA construct transformation and molecular verification
DNA constructs submitted by the customer will be transformed into the appropriate Agrobacterium strain, the plasmid will be extracted and digested with restriction enzymes. The digestion map will be compared with that provided by the customer to determine the stability of the construct.
Cloning service
The molecular cloning services offer over-expression or CRISPR editing with up to 2 guide RNAs. Our process involves assembling DNA fragments into our preferred vector backbone. The resulting constructs are custom designed specifically for integration into the CBL's transformation service, ensuring compatibility and optimal performance.
CBL Prices for 2023-2024:
Customer type | DNA construct transfer into Agro strain1 | Transgenic B1042 | B104 transformation combo3 | Cloning Service4 | B104 Ear5 | Consultation (per hour) |
---|---|---|---|---|---|---|
Internal | 204 | 2400 | 2604 | 811 | 34 | 40 |
External/public | n/a | n/a | 4893 | 1075 | 58 | 82 |
External/private | n/a | n/a | 7050 | n/a | 79 | 124 |
1Price is per construct.
Customer supplies construct DNA and sequence map. Service includes DNA transfer into Agrobacterium, DNA mini-prep and restriction enzyme digestion to verify the transformed Agrobacterium strain.
2Price is for delivering rooted transgenic plantlets ready for transplanting into soil. For each project, a fixed number of explant materials are infected with Agrobacterium carrying the customer construct. An average of 10 independent transgenic events (or lines) will be delivered. Transgenic "event" refers to a transgenic line that has one or more transgenes integrated into the maize genome.
3Price includes both DNA-Agro transfer and transgenic B104 plantlets. See above notes 1 and 2 for details.
4Includes simple gene expression or gene editing with up to 2 guide RNAs cloning into our preferred backbones for Agrobacterium transformation.
5Price is for one intact ear with embryo sizes between 1.5 to 2 mm. Subject to availability unless arrangements are made in advance.
Delivery of transgenic plantlets:
- For internal customers: arrange pick-up with staff by email.
- For external customers: apply for an APHIS BRS interstate movement permit; provide a FedEx account number and full shipping address.
We strive to provide the best possible products and services that we can achieve. However, unforeseen circumstances that are beyond our control may prevent us from achieving a desired outcome. For example, a construct may contain genes that interfere with plant regeneration. Unfortunately, we cannot know that such a construct exists until several steps of the project have been completed. If the staff determines that a transgenic event cannot be obtained, the project will be terminated after consultation with the customer and a fee will be assessed based on the percentage of work completed.
Please contact Jessica Ji for questions or to place orders.