Thank you guys for this really useful forum. I have found some really good advice here. I have some basic questions about intracellular solutions.
1) When making intracellular solution how careful should you be in measuring out salts, some like EGTA are in such tiny amounts that you cant possibly weigh them accurately even for a large volume like 100 ml.
2) I left my intracellular solution at room temp for 2 days. It has a drug (QX314) in it, should it be ok, or I need to trash it?
3) is it ok to use millipore water or do i need to use RNAse and DNase free water.
4) What is the best way to adjust pH. I have noticed that pH changes substantially with increasing volume. I make 140mM CsCl solution. The pH is usually was way off 7.3. I was using ~300 mM CsOH to adjust it, and ended up adding a good volume. Maybe ~4-5 ml. Is it ok to have a 5-10% error margin in volume, if the osmolarity is in the right range?
5) Is a 270 mOsm osmolarity ok?
There are so many of these small questions. is there a good practical guide that beginner's like me can refer to?
Would really appreciate any advice.