I also spent a fair amount of time researching safety protocol for sarkosyl. In higher concentrations, sarkosyl is a particularly nasty substance. The stock solution that was purchased for me is a fairly strong solution: 30%. As noted in the previous post, the "recipe" for the buffer calls for 25g of sarkosyl and as I mentioned my stock is a 30% solution. This was done intentionally, the powdered form of sarkosyl is deadly upon inhalation. Using sarkosyl in solution greatly reduces the risks associated with its handling. It does make the calculations of how much to add to get the appropriate concentration in the final solution a little more complicated, but what's a little extra math for safety's sake. While researching handling safety of chemicals is generally a good lab practice, we are being especially cautious with sarkosyl as it is a cell lysis buffer - it breaks down cell membranes, and I would like to keep mine intact.
Protocol for handling will include the usual necessary PPE: gloves (I did research solubility factors to select appropriate gloves), safety goggles, and lab coat to prevent accidental contact. I will also be handling the stock solution in the fume hood to avoid any vapor contact. The buffer I am creating will be much easier to handle as the sarkosyl will be diluted down to 5%.
I have also scaled back the overall volumes of the solutions made. For instance, the "recipe"for the 5% sarkosyl component of the lysis buffer will make 500mL. I don't need to have that much of the buffer, especially as it is untried as of yet. I have scaled back the amounts by a factor of 5, resulting in a total of 100mL. The same is being done for all of the reagents I will be using.
Looks like I am finally about to pick up where I left off at the end of last semester.
No comments:
Post a Comment