Our control experiments (Figure 1) using defined mixtures of cDNA transformants show that SPI can yield data sets in which real positives overwhelmingly dominate the readout.
The availability of these constructs made it possible to generate an approximately uniform mixture of corresponding transformants without induction, and then to test the impact of their expression when galactose was added.
To identify cDNAs which affect growth upon deliberate overexpression, we have cultured samples of the full mixture of 5885 transformants in galactose medium for 30 generations at 30°C and compared their cDNA spectra to that of cells cultured in glucose medium-Figure 2D.
Unexpectedly, increasing the amount of this mixture did not increase the number of transformants per plate in a dose-dependent manner because the transformation frequency was reduced (Table 4).
For the initial selection of transformants, the DNA/cell mixture was applied to BG11 agar plates.
Ligation mixtures were incubated overnight at 14°C to obtain the maximum number of transformants.
Chen, H., Nelson, R. S. & Sherwood, J. L. Enhanced recovery of transformants of Agrobacterium tumefaciens after freeze-thaw transformation and drug selection.
(e) Colony PCR of transformants using primers that target cmx.
The plasmid complement of transformants was verified by PCR and double restriction digestion.
Molecular and histochemical characterization of transformants revealed the stable integration of the transferred genes into the banana genome.
