I am a masters student from Malaysia and I'm currently researching on the
genome of Pseudomnas sp. USM 4-55. However, I've problems performing
triparental mating between E. coli DH5 alpha lambda pir (donor strain), E.
coli DH5 alpha (helper strain) with pRK2073 and Pseudomonas USM 4-55.
My questions are as follows
1. What is the best method of performing triparental mating besides filter
2. What is the optimised duration of time for mating to occur?
I also conduct biparental mating. But it seems that the plasmid (pUTlacZ1)
could not be transformed into E. coli S 17-1 lambda pir by using TSS.
Can anyone tell me how to transform the plasmid into E. coli S 17-1 lambda