Which Roche systems do you have? We have capillary LightCycler 2.0 and plate LightCycler 480. I love them both but 480 perhaps more... Haven't got the chance to try HRM yet... So far only I use them in our lab so I treat them as if I would treat my baby (if I had one) I never used AB but I had the chance to work on Eppendorf realplex (and I still prefer Roche).
There are many variations in buffer composition. Have you tried adding BSA?
And yes, I'm also looking forwards to your replies

That's good news Optimization of the buffer must have taken you some time and its's good to hear that it works. May I ask you what real-time system do you use? We use two Roche systems.

lol Watch out for you lab notes - someone may steal your buffer recipe
I found two articles about homemade real-time PCR buffers but my experience says that if you have slight variation in components between batches of buffer you may get slightly different results. So for now I prefer to purchase commercial reagents. As long as my prof doesn't care about the price I'm not bothered

Yeah, I guess this is not bad. At least I like this kind of work although research should be linked with industry and not only carried out for the sake of knowledge itself. I also try to follow that... Currently I rely on commercial buffer. I tried to find recipes for homemade buffers that would be cost effective and work well, but after days of trying to convince my supplier to show me the exact content of the buffer I partly gave up.

real-time - I like that. Most of my recent work is based on it. I'm finishing toxin genes profiling of my bacteria and then only dendrograms based on RAPDs are left...
Good to hear from you again

I get you Butters, I also have holiday limit - 21 days off (but these don't include state holidays), so it's much better than 9... I guess you can't do anything about it

Yeah, I want more days off... It seems like I switched my metabolism to resting mode You should have holidays as well.

Hi Butters :-) I hope you're doing great. And your research too. I'm lazy cause I've just come back to work from my 10 days holidays and can't seem to start. And there's nobody in the lab to motivate me ;-)

Hi :-) Good to see you're here again. You must have got rather busy time at work. Good luck!

Butters, gotta catch up in the arcade You mentioned something about no challenge there. So I decided to make it more challenging for you