General Requirements

In order to carry out your transformation requests there are several requirements that need to be taken into consideration.

Constructs

• The transformation protocols followed will use Agrobacterium mediated transformation techniques, and the vector provided should be designed for this purpose e.g. contain left and right border sequences.

• Each crop requires a different selectable marker for efficient selection of transformants.

 

• Brassica: 35S Kan

• Barley: 35S Hyg

• Wheat: Ubi Bar

 

We have available a series of constructs containing these selectable markers and details can be found in the 'constructs available' section of this website.

• Constructs for wheat can also be introduced by bombardment and we will require 30µl DNA at a concentration of 1µg/µl.

If you require any assistance with construct design then please get in touch.

 

Please do not send us vectors for transformation without first contacting us.

 

Agrobacterium

• You will need to provide the vector for Agrobacterium mediated transformation in a specific Agrobacterium strain depending on the crop to be transformed.

 

• Brassica:  LBA4404 or AGL1

• Barley:  AGL0 or AGL1

• Wheat:  AGL1

Primers

• We will require a set of primers, associated PCR programme and a gel picture of PCR product to test for the presence of the construct in the Agrobacterium (or DNA if to be used for wheat bombardment) provided. We will also test for the required selection gene.

Time

• The approximate time scales for generating transgenic plants from the Agrobacterium or DNA supplied are as follows.

 

• Brassica:  12 weeks

• Barley:  14 weeks

• Wheat:  12 weeks