Hello everyone.
I'm a master student. I have got a new project about phytase producing bacteria. It is new for me and I have a counterstaining protocol to prove clear zone of isolated bacteria. But, I cannot prepare counterstaining solution. So, Please anyone recommend me what should I do.
Counterstaining protocol that I have.
- add 2% cobalt chloride on PSM for 5 min. Then, removed and replace with coloring reagent consist of 6.25% aqueous molybdate and 0.42% ammonium vanadate. Finally, removed the reagent.
I have three questions.
1. What chemical should I use to prepare aqueous , ammonium molybdate or sodium molybdate or any ?
2. What should I use to prepare ammonium vanadate solution , ammonium metavanadate , ammonium orthovanadate or ammonium hexavanadate ?
3. how much do I add 2% cobalt chloride on PSM agar ?
Thank you for all answers.
Thank you for your recommendation, Phil Geis.
I have already read it. But the counterstaining technique was not used for this research.
Dear Napkhwan,
You will find all the relevant information in my doctorate thesis entitle: "Phytate mineralization by native soil bacteria and phytase overexpressing rhizobacteria in relation to P availability for plants" find the link below.
http://shodhganga.inflibnet.ac.in/handle/10603/72057
In my doctorate work, I tried the above method but could not work proper, my suggestion is plating should be followed by phytase assay. The zone of clearance is due to organic acid production which give false result but representation by BaCl2.
Hope that help you...good luck..
Thank you for your kind answer, Dr. Kuldeep. I'm reading your perfect thesis.
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