1. What is an EZ-Tn5 transposome?
  2. Does the EZ-Tn5 transposome system use a suicide plasmid to deliver the transposon to the chromosomal DNA of living cells?
  3. Can I use chemically competent cells with Transposomes?
  4. Has my bacteria species of interest been mutagenized by EZ-Tn5 Transposomes? Do the Transposomes function well in Gram-positive bacteria?
  5. What are the best conditions to use to electroporate the Transposome into my bacterium?
  6. Can I use this kit for mutagenesis of genomic DNA in vitro?
  7. Can I get the transposon to precisely jump back out of my DNA once it has hopped in?
  8. The bacterium I want to make a knockout library in is already resistant to the antibiotics you have resistance markers for in your standard transposon offerings. What can I do?
  9. Is there any way to scale back on the amount of transposase I need?
  10. Can Transposomes be used with filamentous fungi, yeast or mammalian cells?
  11. I transformed my bacterium but I only got a few drug-resistant colonies. I expected more than what I got. What happened?
  12. Can I use the EZ-Tn5 TET-1 Insertion kit to generate a Transposome for in vivo use?
  13. I’m just getting ready to use the Transposomes for the first time. Are there any particular things I have to watch out for when I do the experiment?
  14. What factors do I need to consider in order to assure a successful transposome mutagenesis experiment?
  15. I’ve got the knockout mutant I’m looking for and I want to sequence the knockout mutant directly. How do I do this?
  16. I performed a marker rescue experiment with your R6Kγ/KAN-2 Transposome, isolated the genomic DNA, digested it and transformed it into my host bacterium (not Epicentre’s EC100D). I got nothing to grow on Kanamycin plates at all. Why?
  17. I did a Transposome knockout experiment in my bacterium, but I am not getting anywhere close to the numbers of resistant bacteria that was published and I’m using the same strain. Is this data real? Why can’t I get the same results?
  18. How many knockout mutants can I expect when I try to mutagenize my particular bacteria?
  19. It takes an awful lot of DNA to transform my particular organism and I’m worried about the cost of the amount of transposon I need to successfully transform my cells, even to get just one mutant. Is there any good way to get around this problem?
  20. Can I make my own EZ-Tn5 transposome with my gene of choice?
  21. Can I sequence my plasmid with this kit?
  22. Can you tell me how the transposon works?