— Ryla Cantergiani 2020/04/10 19:46

LB (originally, Lysogeny Broth, now often called Luria Broth or Luria Bertani) is a liquid medium for growing bacteria. You may use LB when pulling bacterial strains from the freezer, overnighting bacterial strains, or making a new stock of OP-50.

When overnighting bacterial, take what you need using a pipet aide and stripette. When making a new OP-50 stock, use an fresh 250 mL bottle of LB.

[Insert image of media here. To do this, take a picture of the reagent on the shelf, save it on to the shared computer 'mccormicklab1>Wiki-Documents and Figures> Pictures for WIki with a file name of “nameofmedia-image.png” . Select “Add images and other files” from above and upload your image. When the Link Settings tab pops up, select “Direct link to original”, “Align the image on the right” and “Medium Version”, then click insert. You may have to mess with the size. ]

Location: Shelf ???

[Insert Image of lab location here. To do this, take the template of the lab, that is in the wiki folder, and add a red dot in the location of the reagent using InkScape. Export the image to the dropbox folder 'mccormicklab1>Wiki-Documents and Figures> Pictures for Wiki….> Reagents' With the file name of “nameofmedia-map.png”. Then, add the image by selecting “Add images and other files” from above and upload your image. When the Link Settings tab pops up, select “Direct link to original”, “Align the image on the right” and “Medium Version”, then click insert. ]

Notes and Warning: LB can be stored at room temperature.

1. Grab the same volume beaker as media you intend to make (1 L LB = 1 L beaker)
2. Note: you can only do this because the media will be autoclaved in bottles, not the beaker
3. Fill beaker with 1/4 of the milliQ water.
4. Add a magnetic stir bar that is 1/3 the width the flask's bottom. Place flask onto the magnetic stirrer/hot plate. Turn on the stirrer with no heat.
5. Add Bacto Tryptone, Bacto Yeast Extract, NaCl, and 50% NaOH solution. Stir until all reagents dissolved.
6. Turn off stirring, and bring the volume up with milliQ.
7. pH to 7.5
8. Pour into bottles. Fill 1 L bottles to 900 mL, use a 250 mL bottle for the rest. Fill 250 L bottles to 200 mL.
9. Autoclave to sterilize

Note: When adding ampicillin, you want to add 100 µg/mL.

1. Grab a flask that is twice the volume than what you will be making (2 L flask for 1 L media).
2. Fill flask with 1/4 of the milliQ water.
3. Add a magnetic stir bar that is 1/3 the width the flask's bottom. Place flask onto the magnetic stirrer/hot plate. Turn on the stirrer with no heat.
4. Add Bacto Tryptone, Bacto Yeast Extract, NaCl, agar, and 50% NaOH solution. Stir until all reagents dissolved.
5. Turn off stirring, and bring the volume up with milliQ.
6. pH to 7.5
7. Autoclave to sterilize
8. Pour plates. Let sit until solidified, then bag and store in 4C refrigerator.

Note: When adding ampicillin, you want to add 100 µg/mL.