Glove Box for Growing Yeast

[Jdl973]

Well this seems to be the week for the brew projects. Today I built a glove box to play with yeast in.

Ingredients

-Old 10 gal fish tank, glass, off of Craigs List = $10
-0.22" Lexan, Lowes, $10 (the trick is to find some of the "scrap" pieces and make them a deal)
- 2 small hinges, Lowes, $3
- 2 x 4" male pvc adapters, $8
- 2 x 4" female pvc adapters, $8
- tube of silicone caulk , $4
- 1 pr gloves, $8

TOTAL COST ABOUT $50

All the parts (blue stuff is the lexan)





The lexan was cut to fit the inside lip of the tank. An access hatch was fitted and cut as well. Dry fit.



The 4" PVC couplers were cut down. This will allow more of your forearm to be in the tank.



Holes were cut for the couplers.



Another dry fitting



Cut a 3/4" strip of lexan and fitted it on the glove side of the lid, this is where the screws will anchor for the door and it will produce and overlap lip to keep the door air tight. Holes were drilled for the screws. You have to do this oR the plastic will split like crazy.



Final dry fit. Almost done. It is too cold (and too late) to silicone everything. Need to clean up everything as well to get a good bond. Not bad for a couple of hours.





Will try to get it finished tomorrow and will post pics.

Jason

[wlockwood]

Nice "clean room".

Back in the day when I worked in a lab we had a UV light mounted on the top of our boxes (inside), and there was room for flame etc..., these were plywood boxes with a clear front (UV filtering). You could just reach in, no gloves but protected from draughts (plus the UV light). I saw a "germ light" at CVS the other day.

[Jdl973]

Yea I was thinking about using a UV to "sterilize" the box before use. Back in my biochem days we used UV light in our hoods to "kill" any DNA before we started work.

J

[Jdl973]

The box is done and I inoculated my first plate. I tried using the box with gloves, but it was just too constrictive. So I use the box sans gloves and it seems to work just fine. Gives me a clean area with no drafts to work in. I also had some issues with the alcohol lamp inside the box...it fogs the glass. So i put it out to the side (on the right) and move the loop in and out.

Yeast growing setup:





First plate after 24 hours. I hope the hazy stuff is yeast



Jason

[Jensen]

Great job on the work box Jason. Let me know when you up-grade, I'll buy that one!! I would think a heat flame inside the box might cause a convection current, actually defeating the purpose... UV baby!! Keep up the good posts-- You and Chris are keeping this place interesting on an otherwise cold January...

[klickcue]

Though I have an alcohol lamp, I just use a propane torch. An alcohol lamp works fine.

The only suggestion that I have is get rid of the air locks on your growing containers. As the yeast produce some CO2, you want the partial pressure of O2 to mix back into the head space. At this stage of the game, you want a lot of yeast growth with O2 and very little CO2 production. Aluminum foil works great.

A pound of CO2 equals a pound of alcohol.

[Jdl973]

Aluminum foil it is. I was wondering about that, thanks for the tip

Yes UV would be the way to go. I got one of those electric toothbrushes for Christmas, it cam with a UV brush cleaner....Hummm. I'm a thinking there might be some "modifications" in its very near future...hmm.

[klickcue]

Jason, it looks like you are on your way to great things. It won't be long and we will have to call you 'The Banker'

[Jdl973]

Yeast at 48 hrs



Can start to see the white inoculation lines. Ain't they cute little buggers (wow I got to get out more )

So far, everything is white, no blue, green or brown meanies.

Jason

[Jdl973]

Yeast at 72 hours. The colonies are now visible threw the glove box. The dish on the right is the "Control" just to make sure nothing is growing in the box.





In a couple of days I will transfer some colonies to 3 slants.

Jason

[klickcue]

Excellent!

Did you use a loop or a needle, it is hard to tell from the photographs?

From here, transfer 3 to 5 single colonies to each slant with a loop. You want an average of the gene pool.

Nicely done.

[Jdl973]

Used a loop, my technique was a bit sloppy and my media was not very smooth (will bloom it next time before heating...again ...duh )

Good idea on snagging several colonies per slant. No swimming in the shallow end of the gene pool please.

Jason

[klickcue]

The blooming of the agar (let the agar set in the cool wort before heating) should help you get a little finer texture as well as stirring it up while it is warming up. Just think that you are making gravy from flour.

It is just like getting dough balls in your mash where you gelatinize the outside layer of the starches and seal the grain into a ball.

I believe that the gel state for agar is around 146 oF , so keep you solution above that temperature and you will pour nice petri dishes and slants.

I am glad that your new agar product is working better for you. The 3 to 4 percent w/v should be correct instead of the 8 percent that you were getting before. You might even try a 2 percent w/v for your plates, it will scratch a little easier. Look for tearing with your needle. If it tears, increase your percentage of agar.

You should see some nice colonies on your dish now that it has been 4 days.

Looks like you are having fun. Keep the beasties happy and they will make you happy.

[Jdl973]

Day 4, the kids grow up so fast (sniff)

96 hours and individual colonies are viable and taking root.





This has been a very fun and rewarding project. It has been a great deal easier than I thought it would be, especially with all the help I got (thanks Chris).

What should I do for my next beer project....

Now to make some beer!

Jason

[klickcue]

I think that I will throw this into this thread to keep Jason's fine work together.

The is an over populated slant of 2565. You can see where I was harvesting yeast from the lower right hand area. The petri dish could have been used as shown to make a great starter for a beer.

This was done with a needle instead of a loop. Notice that the edges are nice and round. An excellent growth of cells. The needle had to much liquid on it and the passes where not drawn out far enough or there would have been a lot more separation between cells. The needle only needs to be dipped into the starter yeast since there will be a lot of yeast on the needle. If the needle had been pulled back through at 90 degrees, the separation would have been better. Still pretty though.