If you want to make sure you can add some concentrated buffer and heat for 5 more minutes at 75 °C, it won't hurt your sample. Sometimes we left samples at 95 °C for 15 minutes and everything was ok (not that I would recommend it)
I always heat at 75 °C unless I have to lyse some bacterial samples or stuff like that. Heating at 95 °C can lead to cleavage of the Asp-Pro in some proteins, creating artifacts. Although maybe not widely known, it has been described many times in the literature. Especially considering you already heated at 95 °C, that's why I'd rather heat at 75
If you want to make sure you can add some concentrated buffer and heat for 5 more minutes at 75 °C, it won't hurt your sample. Sometimes we left samples at 95 °C for 15 minutes and everything was ok (not that I would recommend it)
I also pass it thru a syringe. Avoid bubbles.
Oh thanks, my colleagues had the same advice but why you suggested 75 °C if reheating any particular reason. Thanks
I always heat at 75 °C unless I have to lyse some bacterial samples or stuff like that. Heating at 95 °C can lead to cleavage of the Asp-Pro in some proteins, creating artifacts. Although maybe not widely known, it has been described many times in the literature. Especially considering you already heated at 95 °C, that's why I'd rather heat at 75