We have introduced TAC clones containing Arabidopsis genomic DNA into A. thaliana, and these experiments have shown that the length of the DNA insert in the TAC vector is not directly related to the efficiency of transformation. We routinely use the vacuum-infiltration protocol for Arabidopsis transformation; tissue culture-based protocols can be used for Arabidopsis mutants that are resistant to this procedure.
Of 36 independent hygromycin-resistant plants obtained with four distinct TAC clones harboring DNA inserts of 40 to 80 kb, 32 plants contained the sacB sequence, indicating that, in most transformants, the entire insert was transferred to the Arabidopsis genome. The integrity of some transgenes was also confirmed by Southern blot analysis of I-SceI digests.