Hi Colleagues,
I am going for transduction of Salmonella Typhimurium. As per the protocol available with me the method goes in the following way:
1. Take 100 ul of overnight grown culture of Salmonella Typhimurium (with choloramphenicol resistance gene) and 100 ul of phage lysate.
2. Incubate at 37 deg C for 1 hour.
3. Plate the culture in a LB Agar plate with Chloremphenicol and 10mM EGTA.
4. Incubate the plates in 37 deg C for overnight.
So, my question is can I replace EGTA with EDTA?
Please guide me.
Thanks for cooperation.
EGTA is used to chelate Ca2+ and prevent phage readsorbtion. EDTA has lesser affinity for Ca2+ and higher affinity for other divalent ions (ie Mg2+) and will inhibit growth. EGTA can be omitted but you must ensure that your clones are not infected (ie streaking clones on green plates)
- Badges
- Science topic
- Similar topics
- Physical Sciences
- Materials Science
- Plating