Need feedback on my lab setup

[Bunker]

Hey guys. I'm going to start brewing my own gear and I've been going through a list of supplies and materials, and I would appreciate some feedback.

I'm not going to cheap out on anything so I'm going for professional setup:

250ml beakers (autoclavable) x 4

50ml beakers (autoclavable) x 4

Autoclave

Magnetic hotplate stirrer

Hepafilter air system for the room

Bottle top filter system with vacuum pump (.2um filters good for all gear??)

Pipet pump 10ml pipetes

Hand crimper

___________

Now I was planning on buying unsterilized vials since I'll have an autoclave and just filling them with a pipet and crimping. I'm gonna autoclave the beakers I'm mixing my solutions in and also the media collection bottle from the filter.
After filtration will exposing the filtered solutions in the media botte to air and transferring with the pipet to the 10ml vials be a concern for air contamination? And can I use the same pipet for the entire transfer of each batch?

 

[Wunderpus]

My honest suggestion would be to do a few brews a simpler way, with sterile vials and a good disposable vacuum filter. Get the hang of it before you invest in shit like an autoclave... Just my thoughts on the matter.

 

[Bunker]

I have most of this equipment already bro. I get it at cost and I'd like to do it as professional as possible. Of coarse I will do smaller batches to get the technique down, but I'm just trying to be overkill when it comes to sterilization.

 

[Johne71]

Yes...filtering all oil into one container is quick and easy, but you are correct. Getting that oil from there to each vial poses a risk for contamination. I've seen electronic pipettes that allow you to aspirate up to 50-60ml at once into a sterile (of course) pipette. Then you can dispense (defined volume) into each auto leaves vial.

I'm really not sure how most move oil from a large filter flask to each vial, but I'm pretty sure they are taking chances.

Without a laminar flownhood are risk is increased,but we work within our limits.

 

[Bunker]

Yes...filtering all oil into one container is quick and easy, but you are correct. Getting that oil from there to each vial poses a risk for contamination. I've seen electronic pipettes that allow you to aspirate up to 50-60ml at once into a sterile (of course) pipette. Then you can dispense (defined volume) into each auto leaves vial.

I'm really not sure how most move oil from a large filter flask to each vial, but I'm pretty sure they are taking chances.

Without a laminar flownhood are risk is increased,but we work within our limits.

That's exactly the problem I'm trying to work on. Now my area will be confined and I'm going to be using a hepafilter system in the room so I hope that will help. I'm just trying to be cautious about the transferring. After filtration and the BA BB added though is there really such an environment for contamination to take hold though?

 

[Johne71]

After filtration and the BA BB added though is there really such an environment for contamination to take h

Not likely. Lint may fall in or something like that, but the BA should keep anything from growing. I'm assuming a lot of people use a 60cc syringe an take as much as they can from the filtered oil and then remove the needle to dispense easily into each vial. Maybe the needle makes it easier. I have gotten presterilized vials.

 

[Bunker]

Not likely. Lint may fall in or something like that, but the BA should keep anything from growing. I'm assuming a lot of people use a 60cc syringe an take as much as they can from the filtered oil and then remove the needle to dispense easily into each vial. Maybe the needle makes it easier. I have gotten presterilized vials.

That's what I figured. I'm pretty confident my room will be filtered and cleaned to excellent standards. I'm also wearing tyvex suite and mask. My major concern was airborne pathogens but I'm convinced I was just over thinking the process.

 

[Johne71]

You are much much more cautious than I have been. Again, I'm filtering directly into my own personal use vials. I'm only making enough to last a few weeks at a time.