After Using a sterile pipette tip or toothpick, select a single colony from your LB agar plate
The use of Shrink-Seals (REF R522600) or gas-permeable tape to secure the plates is recommended to prevent exogenous contamination, avoid moisture loss, and protect personnel from exposure to pathogenic molds
This Thermo Scientific Chemicals brand product was originally part of LB-kanamycin agar plates
Plates will keep refrigerated for 4-6 weeks
0 g agar-agar 1 L Sterile H 2 O Sterile plates of your desired size - we usually use 60 mm x
How to make a 25-50 mg/ml Chloramphenicol Stock Solution
Chloramphenicol – add 1ml chloramphenicol stock (at 25mg/ml) per liter of agar to obtain a final concentration of 100ug/ml
In most cases you will add 1 ml of stock antibiotic per liter of solution
Add about 800mL of dH O and dissolve the big chunks and stir with a magnetic bar until it's
Pour hot/ warm (melted) media at 20ml per petri dish [Note: stack plates on top of each other when preparing several plates to preven plates from sweating] PREPARATION OF LB-AMP PLATE (500ml) Add 12
Thaw and mix (stir) 100 mM stock solutions of dGTP, dATP, dTTP, dCTP (NEB; in freezer) Master Mix: 60 μl sterile dH 2 O
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Then, filter sterilize solutions by pushing them through a 50 mL syringe fitted with a 0
In general, use carbenicillin for ampicillin selection Plate 50 µL of transformed E
Ampicillin (Amp/A): *Working concentration 100 ug/mL freezer stock 100 mg/mL (in water) Add 1 mL of 100 mg/mL ampicillin (dissolved in water) into 1
Note: If desired, 20 units of Penicillin and 40 µg Streptomycin per ml of the medium may be added to make the medium selective for fungi
North American Regional Complaint Center 1-844-8BD-LIFE (1-844-823-5433)
For 500 milliliters (ml) of agar mixture, you can fill 25 average 1
Cool to 55°C
Prepare the desired volume of stock solution by dissolving chloramphenicol (Sigma-Aldrich C1919) at 10 mg/mL (or 20 mg/mL) in absolute ethanol
Add the appropriate amount of desired antibiotic to the solution (500µL if you are using a 1,000x antibiotic stock) and swirl to mix
For use, add 5 mL of the antibiotic mix to 1 L of Chloramphenicol (methanol) 10 20 10 40 80 0
2
I reduced the chloramphenicol concentration to an intermediate concentration of 68 ug/ml (about 2
To process the specimen, stream the specimen being studied on the medium using a sterile inoculating loop
Add Water and Stir until clumps are gone
You may need to tilt the plate slightly to get the agar to Principle of Sabouraud Dextrose Agar (SDA) Sabouraud Dextrose Agar contains digests of animal tissues (peptones) which provide a nutritious source of amino acids and nitrogenous compounds for the growth of fungi and yeasts
5 ug/mL final concentration) 2
Pour ~20mL per 10cm
6
Add 40 μL of the Thermo Scientific X-Gal Solution (20 mg/mL You'll get a detailed solution from a subject matter expert that helps you learn core concepts
10 μl dGTP
Sabouraud dextrose agar is a selective medium for it is used to isolate yeast and fungi and at the same time permit the growth of filamentous bacteria like Nocardia
Potato Dextrose Agar (PDA) P otato Dextrose Agar (PDA) is a common fungal growth media made from an infusion of potato and dextrose