Bacterial media
Bacterial media kdorfman Thu, 01/17/2013 - 18:53Freezing bacteria
Freezing bacteria kdorfman Sat, 12/05/2015 - 18:44To Freeze:
Pick a new colony, grow overnight in LB
inoculate LB from the overnight culture
Put 0.15 mL glycerol into sterile cryovial.
Add 0.85 mL mid-log culture (OD =~0.4). Pipette up and down to mix thoroughly
Freeze. Store in -80 if possible.
To thaw:
Scrape surface of frozen stock with sterile stick
Streak on agar plate
LB Broth
LB Broth kdorfman Fri, 09/25/2020 - 17:54LB broth from mix
LB broth from mix kdorfman Tue, 05/28/2013 - 17:48LB Broth Miller Luria-Bertani
Fisher DF0446-17-3 500 g $40.61.
25 g/L
LB from scratch
LB from scratch kdorfman Fri, 09/25/2020 - 17:38LB (Miller) from scratch
LB agar
LB agar kdorfman Thu, 01/17/2013 - 18:55LB agar granules
LB agar granules kdorfman Tue, 06/25/2019 - 20:01Make with granulated LB (Miller) agar, Fisher BP9724
LB agar powder
LB agar powder kdorfman Tue, 06/25/2019 - 20:00Make with LB (Miller) Agar powder, Difco 244510
LB broth + agar
LB broth + agar kdorfman Tue, 06/25/2019 - 20:0025 g LB + 15 g agar /liter
LB antibiotic
LB antibiotic kdorfman Wed, 10/01/2014 - 16:34Table of antibiotic stability in poured agar
line | antibiotic |
---|---|
black | Carbenicillin (ampicillin) |
green | IPTG |
LB-sugars
LB-sugars kdorfman Wed, 10/01/2014 - 16:19LB + sugars for Lac-operon work
sugar (or analog) | MW |
---|---|
mono saccharide | 180.2 |
disaccharide | 342.3 |
IPTG | 238.3 |
glucose, galactose, or fructose
Lactose is not very soluble in cold water.. Boil water, then set it to stir on a stir plate. Gradually tap in the lactose powder.
lactose, maltose, sucrose
IPTG
Filter sterilize
Add 50 µg/mL Kanamycin as indicated here.
MacConkey agar
MacConkey agar kdorfman Thu, 01/17/2013 - 19:41MacConkey Agar
- Fisher 212122 2 kg (Difco)
- Krackeler 10-211387 500g (via Sigma)
- HiMedia MacConkey agar, granulated GM081 (Fisher NC1876763, VWR 95020-204)
to distinguish Lac+ and Lac- bacterial strains.
50 g/L
leave stir bar in
put bottles on stir plate near sterile hood until handle-able
If you accidentally buy
Min med for GFP cells
Min med for GFP cells kdorfman Tue, 10/23/2018 - 15:53From https://wahoo.cns.umass.edu/node/857
For SfGFP cells, try M9 + 0.2% glycerol + 0.01 mM FeSO4
Mueller Hinton agar
Mueller Hinton agar kdorfman Thu, 01/17/2013 - 20:0590922 Mueller Hinton Broth 2 from Sigma
22 grams per liter, consisting of:
Casein acid hydrolysate 17.5
Beef extract 3.0
Starch 1.5
Final pH 7.3 +/- 0.2 at 25°C
P-Glo Bacterial media
P-Glo Bacterial media kdorfman Mon, 08/26/2019 - 22:26Make 60 mm plates
plain LB plates
LB/Amp (or Carb) plates (1 black stripe)
LB/Amp/Ara plates (1 black, 1 red stripe)
2019 - for QBoC:
- transformed pglo plasmid (1660405EDU) from the BioRad kit into
- DH5 alpha E. coli (Invitrogen 18265-017) from the -80
- Lyophilized cells (BioRad 1660408EDU) did not arrive in time
Used the Invitrogen protocol.
Plated on:
- LB
- LB amp
- LB amp ara
Saw gfp colonies on the arabinose plates. Collected, grew up in LB amp, froze in 15% glycerol.
LB-Amp-Ara
LB-Amp-Ara kdorfman Mon, 08/26/2019 - 22:27LB with Ampicilin or Carbenicilin and arabinose
Arabinose stock 200mg/mL in water, sterilized by filtration (=100 X)
Final concentration = 2 mg/mL
- Arabinose MW = 150.13
- 2 mg/mL = 2 g/L x 1 mol/150.13 g = 0.013M = 13 mM
Carbenicilin stock is 1000X
minimal medium (glycerol)
minimal medium (glycerol) bcrcstaff Wed, 11/14/2018 - 17:34For motility strains