Oct 24, 2018
Making Luria Broth (LB)
- 1BosLab
- BosLab
Protocol Citation: Angela N Kaczmarczyk 2018. Making Luria Broth (LB). protocols.io https://dx.doi.org/10.17504/protocols.io.i6gchbw
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: July 30, 2017
Last Modified: August 01, 2019
Protocol Integer ID: 7080
Abstract
Luria Broth (LB) is a nutrient-rich liquid media that is used to grow bacterial cultures. Luria Broth consists of Tryptone, Yeast extract and Sodium Chloride mixed into distilled water.
Guidelines
Equipment needed:
- 500 mL or 1-liter bottle
- graduated cylinder (100mL or greater)
- scale for weighing
- Autoclave
- autoclave gloves
Materials
MATERIALS
Distilled Water
Luria broth powder
Safety warnings
The LB powder is very fine so handle it slowly and with care. The dust can get all over the bench and the powder may irritate your lungs if inhaled.
Before start
Give yourself about 2.5 hours to finish making the sterilized, ready-to-use LB broth. Making the LB broth doesn't take long, but the autoclave use can take up to 2.5 hours.
Read the instructions on the LB container to figure out how many grams per liter are needed to make liquid media.
Take the sterile spoon and spoon LB onto weigh tray. The LB powder is very fine so handle it slowly and with care. The dust can get all over the bench and the powder may irritate your lungs if inhaled.
Note
The LB powder is very fine so handle it slowly and with care. The dust can get all over the bench and the powder may irritate your lungs if inhaled.
Pinch tray corners and carefully pour the powder into bottle.
Measure 100 mL (or desired volume of LB broth) of distilled water in a graduated cylinder. Add the water to the bottle and close it.
Shake media bottle vigorously. Loosen the cap so that it is almost falling off.
Secure it with a piece of autoclave tape and label it with your name, date, and “LB broth” on non-autoclave tape and stick to the front of the bottle.
Put something at the bottom of autoclave basket so you don’t submerge your autoclave bottles.
Place in autoclave basket. Autoclave when ready. See autoclave instructions.
When autoclave is done remove from autoclave per autoclave instructions. Twist cap closed to seal LB. Place in autoclaved media bin with label, date, name.