Making Bottles and Media Splitters

While some groups like to use large carboys for media, I find that individual bottles makes media preparation easier. It’s also essential for experiments with many different media types. Here’s how I make media bottles from standard 1L lab bottles.

You’ll need:
2 Female Luer Lock w/ 5/32" barb, polypropylene
1 Male Luer Lock w/ 5/32" barb, polypropylene
1 Luer Cap
~ 1 ft of silicone tubing
1L borosilicate glass bottle or similar

Drill with 3/16" bit


First, drill a hole directly down through the cap, be sure to have something safe to drill on the other side.

Next, position one of the female luers barb side up, and push the bard through the top of the cap so the luer side is facing up. This takes a little force, so I like to use something with a hole in it to help prevent damage if I slip.

Next, cut two pieces of tubing. One is for above the cap, and can be as short as 3". Put the two remaining luer adapters on this piece. This flexible piece makes it a little nicer to connect lines to bottles. The other should be at least 10" so that it makes it from the barb on the cap all the way to the bottom of the bottle.

The long straw should reach the bottom of the bottle and bend slightly. If it’s too long it will be annoying to screw the bottle cap on, and might get stuck against the wall. If its too short, it won’t be able to reach media at the bottom of the bottle. Your bottles may vary: for our 2L bottles, we need a 13" straw.

Rinse out the straw with diH2O, and add a mini luer cap to the top of the straw before autoclaving, and you’re good to go!

You can also make splitters with the same tubing and similar luer attachments, allowing you to split one media bottle to multiple input lines. We usually sterilize the splitters using 10% bleach and ethanol (same as the lines on eVOLVER), but everything I’ve shown here is autoclavable.


What are the McMaster-Carr catalogue numbers/names for the matching luer 3-way splitter/attachment pieces? Are there 4-way attachment pieces? Thanks!

Here’s a list of many barbed connectors

And these work with the 5/32" silicone tubing? (Just want to double check since the diameters and compatible tubing materials don’t match in the item’s description.)

Ah my bad. Yes get the appropriate one. Sorry about that.

I will note that since the McMaster barbs are much larger than the Nordson barbs, even though the tubing is 5/32" it might be preferable to order the smaller 1/8" size to make it easier to assemble. Haven’t tried this however.

Ok, good to know, I haven’t had trouble connecting the McMaster-Carr 5/32" plugs and sockets to the silicone tubing. I found these 53055K479 connectors; we might try these first since they are compatible with 5/32" silicone tubing and can be autoclaved (the only other ones McMaster-Carr has of this specification cannot be autoclaved).

Hi all,
I am a bit confused with the list of parts available on the Jove paper for making media bottles and connectors.
The tubing is ID 1/8"
The T for media input branching is for tubing of ID 1/8"
Then for the quick connectors to build the media bottles, there are two identical sets of male and female luer locks in 1/16" and 5/32".

So with 1/8" tubing, using 5/32" luer locks seems good (but not sure then why the T is not 5/32" as well). But what are the 1/16" luer lock for ? Why two different sets, both indicated as dedicated to media bottles, in 1/16" and 5/32" ?

I am probably missing something but some clarification would be great ! Thanks !!!

1/16" leur locks are for the smaller silicone tubing (e.g. ones connected directly to the peristaltic pump) or lines that go into individual vials. Just good to have extra ones of those.

The 5/32" connectors are for the larger tubing as seen above.

Does that help? It doesn’t really matter what precise part/ measurements they are as long as they fit the tubing you are trying to interface with the system.

Thanks for the info ! Order is through.