Biology newbie in need of help regarding autoclaving

[azzurrifan]

As the title says this is my first foray into biological lab techniques. I'm actually a chemist but my lab has started some biological chemistry work. WE'll be using peptides to synthesize inorganic materials basically. What I want to know is for this kind of work is it necessary to autoclave everything you use and when you autoclave do you have to autoclave all your equipment (including things like the bottles that contain buffer)?

[Helen Troilo]

That entirely depends what you are working with. For example if you are supplied the proteins and they are frozen/in powder form and you resuspend them before use then there is no need to autoclave anything. If you are growing the proteins up from bacteria then pretty much everything involved with the bacteria needs to be sterilish. Buffers for protein purification would probably have a longer shelf-life if you autoclave them, but its not worth it as the buffer components would likely start to precipitate out long before any bugs move in.

Perhaps if you could tell us more specificially what you're doing we could answer better?

[azzurrifan]

The peptides have been synthesized for us and are in powder form. THey need only be dissolved in buffer. Basically we will be using the peptides to reduce metal ions and form metal nanoparticles. The peptides would then serve as a stabilizing agent.

[admin]

Hi Azzurrifan,
generally in molecular biology labs we autoclave most things that can be autoclaved - this includes pipette tips, equipment such as glassware, and even buffers and water in glassware.

The reason being if you spend X amount of time and money, you do not want to lose them both because you didn't want to spend a bit of time autoclaving. Autoclaving only costs time, not money usually. Even better, get a bunch of tips, glassware and buffers all at once - autoclave them - and they can last you your whole project probably.

Some people cut corners, but you have to assess where you can cut them. I would mainly keep on the safe side and work with the peptide sterile (using autoclaved stuff), because the last thing you want to find is that you have contamination in your stocks - or your amazing findings are actually due to contamination - or worse that they your findings may be lost or be complicated by things growing in them.

I think it is important to also find out if autoclaving will affect your experiments negatively, does it generate nanoparticles that may compromise your study? I am not sure about this but would pay to look it up - what do others in your field do?