One milliliter of whole mycelia extracts from the TU-6 or TU6-vgb+ transformants was treated with 20 mg of sodium sulfite.
The transformants were treated with MMS and cell extracts were prepared at different time points.
The transformants were treated with 0.01% MMS for 0, 1, 2, 3 and 4 h, and the survival rates were measured by the colony formation assay.
In this screening platform, three different yeast pools were interrogated with a single TAG4 array: a homozygous deletion pool, a heterozygous deletion pool, and a pool of genomic library transformants, each treated with a compound of interest.
Subsequent to transformation process and selecting transformants (as single colonies on LB broth containing ampicillin, as an antibiotic resistance marker), the plasmid was extracted from transformants E. coli and treated with double digesting using BamHI and NdeI to confirm the successful transformation.
Primary Arabidopsis J1551 UASGAL4 AtHKT1 1 transformants grown in soil and treated with a low concentration of NaCl (2 mM) showed 20% lower leaf Na (Figure 1D) and significantly higher K, P and Zn (averaged over all independent T1 lines produced) than J1551 (Table S1).
We report here that treatment with LiAc and DDT before electroporation increased transformation efficiency to 225 ± 52.5 × 107 transformants per μg DNA, while with untreated cells or treated with LiAc alone transformation efficiency approximated 1.2 ± 0.5 × 105 transformants per μg DNA.
XhoI-digested and EcoR1-digested DNA from transformant R. montanensis cells was treated with the DNA TerminatorR End Repair Kit, ligated into the pSMART-LCKan vector with Clone Smart DNA ligase and electroporated into E. cloni™ 10 G electrocompetent cells according to the manufacturer's suggested protocols (Lucigen, Madison, WI).
Mouse WR19L transformants expressing Fas (W3 cells) were treated with Fas ligand (FasL) for 30 min, washed, and then further incubated for 60 min. Following FasL treatment, more than 90% of the W3 cells were Annexin V positive, and only small percentage were positive for both Annexin V and propidium iodide (PI), indicating that the majority of cells had undergone apoptosis but not necrosis.
Transformants containing an integrated vector were then treated with 5-FOA to select for excision of the plasmid sequences leaving the EI expression cassette in the chromosome.
