Is there any difference in using doxycycline hydrochloride or doxycycline hyclate salt for doxycycline-inducible gene expression?Any special requirements for doxycycline reagents used for gene expression induction, or can I just use any doxycycline antibiotic, suitable for cell cultures? Please share any personal experience and tips.
Hi Marija,
I have always used Doxycycline hyclate dissolved in distilled and sterile filtered water. You can actually filter the solution with .2 um filters after you've prepared it. We used to prepare 2 mg/ml stocks and store them at -20 to be used at 1:2000 dilution when needed. Working concentration for gene induction are usually between 10 and 1000 ng/ml. Inducible gene activation can be observed from 6-12 hrs with maximum expression reached at 48-72 hrs. We used to refresh Doxycyline in cell culture every 2 days. After thawing, the stock vials can be stored for 1-2 weeks at 4 C away from light.
Hope it helps
Francesco
Hello
they both have the same active form but the hyclate is just more soluble in water.
- Badges
- Science topic
- Similar topics
- Biological Science
- Genetics
- Gene Expression