It is not a good idea to guess on a solution. Things could get painful or expensive quickly. You really should start with some standard/common triage techniques and make an educated decision.
Standard process for troubleshooting performance problems on a data driven app go like this:
- Review DB indexes (unlikely) and tune as needed.
- Check resource utilization: CPU, RAM. If your CPU is maxed-out, then consider adding/upgrading CPU or optimize code or split your tiers. If your RAM is maxed-out, then consider adding RAM or split your tiers. I realize that you just bought new hardware, but you also changed OS and IIS. So, all bets are off. Take the 10 minutes to confirm that you have enough CPU and RAM, so you can confidently eliminate those from the list.
- Check HDD usage: if your queue length goes above 1 very often (more than once per 10 seconds), upgrade disk bandwidth or scale-out your disk (RAID, multiple MDF/LDFs, DB partitioning). Check this on each MySql box.
- Check network bandwidth (very unlikely, but check it anyway)
- Code: a) Consider upgrading to .net 3.5 (or above). It was designed for better scalability and has much better options for caching. b) Use newer/improved caching. c) pick through the code for harsh queries and DB usage. I have had really good experiences with RedGate Ants, but equiv. products work good too.
And then things get more specific to your architecture, code and platform.
There are also some locking mechanisms for the Application variable, but they are rarely the cause of lockups.
You might want to keep an eye on your pool recycle statistics. If you have a memory leak (or connection leak, etc) IIS might seem to freeze when the pool tops-out and restarts.