Hello everyone!
I transformed my vector PL4440 containing the gene of interest, into HT115 competent cells. However the culrure can not grow on LB agar plates (amp + tetr ) antibiotic. Culture can grow from liquid to liquid LB containing Ampicilin 50 mg/ml and tetracycline 12,5 mg/ml. Had anyone this experience ?
Sometimes when you start combining antibiotics the cumulative effect gets to be too much, although usually it works fine. However you really don't need to maintain selection on Tet, that is a chromosomal insertion so is stable. You can just select on amp plates for your plasmid and test every once in a while to be sure you still have the Tet marker.
My HT115 with pJET growed ok in plates with that same concentration of antibiotics. It did grow much slower though. How are your competent cells and transformation protocols?
-->I am not sure where you got your HT115 colony but I would encourage you to keep the Tet, if your source is not 100 % sure (this is if someone donate it to you instead of buying it), until you get your glicerol stocks.
- Badges
- Science topic
- Similar topics
- Engineering
- Materials Engineering
- Powders