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
Scissors/razor
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.
