Table of Contents
Making an OP-50 Stock
— Ryla Cantergiani 2020/04/10 21:42
Overview
We use OP-50 stocks for seeding NGM plates. Unfortunately, while LB media is great for growing up OP-50, it's also great for growing up all sorta of other contaminants as well. If you see gross floaties in the stock, or suspect your stock is contaminated, you'll need to make a new stock.
In this section you put why we do this protocol and how it works very generally, as well as what the protocol(s) immediately preceding this and immediately following this are (with wiki links), if applicable.
Materials
| Material | Amount |
|---|---|
| pipette tips | 1 per stock |
| frozen OP-50 | 1 aliquot |
| LB (200 mL) | 1 250 mL bottle per stock |
Procedure
- Ethanol and wipe down your benchtop. Turn on a bunsen burner.
- Grab an unused 250 mL bottle of LB. Label it “OP-50 in LB” with your initials and the date.
- Retrieve an an aliquot of OP-50 from the -80C freezer (MM158-B).
- Using a pipette tip, scoop out some of the OP-50 and put it into the LB. Swirl the LB to mix.
- Seal the bottle and place on the shelf to grow up. To speed up growth, you can also place the bottle in the 37C incubator.
- if using the 37C incubator, don't fully seal the bottle.