Hi everyone,
I would like to add some guanine, cytosine and adenine, independently, to cell culture. I can't dissolve them in DMSO or in water, even turning up the temperature until 95°C. Only cytosine dissolve at 55°C but precipitate again when the temperature goes down. On the specification sheets of the products, it is written they are soluble in HCl.
- HCl 0.5M for cytosine up to 100mM
- HCl 0.5M for adenine up to 20mg/mL
- HCl 5M for guanine up to 25mg/mL
At the concentrations I would like to use them on the cells, even trying to prepare these bases at a 1000X concentration, two problems come to me:
- I'm not even sure they can dissolve in the appropriate volume for the 1000X concentration: the mass/volume being higher than the specifications
- I tried to dilute, even the lowest HCl concentration (0.5M), a thousand time in my cell culture medium and measured the pH with a pH paper --> pH came down to 6. This is not suitable for cell culture.
Do you have any idea how to dissolve these bases?
Carole Trzaska When you dissolve dNTP in HCl, it would be acidic which may harm your cells. why dont you try with TE buffer to dissolve, just take a few microlitres of the dNTP and give a try .
http://cshprotocols.cshlp.org/content/2009/2/pdb.rec11661.full?text_only=true
All the Best.
Thank you for your reply, I'm actually not working on that anymore. However, it was directly from the dNTP in powders, so I had to dissolve them first.
Adenine can be dissolved in water by stirring at 60 0C and remains long after the temperature decreased to RT.
Cytosine dissolves in water by stirring at RT.
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