Cloning & Expression

S-Gal Preparation


Simplify Preparation with S-Gal™/LB Agar Blend


S-Gal/LB Agar Blend X-gal Time Saved





Prepare antibiotic stock solution and filter-sterilize.


Note: Powdered S-Gal/LB Agar blend is stored at room temperature.



Time: 15 min.
Prepare X-gal stock solution in dimethyl formamide.

Prepare aqueous IPTG and filter-sterilize.

Prepare antibiotic stock solution and filter-sterilize.


Note: Stocks not used immediately require refrigeration or freezing. X-gal is light sensitive; stock solution should be protected from light.

Time: 45 min.
30 min.
Mix mix S-Gal/LB Agar Blend in 500 ml deionized water.

Autoclave at 125°C, 15 psi, for 20 min., using slow exhaust cycle.


Time: 45-60 min.
Mix nutrient agar medium in deionized water.


Autoclave at 125°C, 15 psi, for 20 min., using slow exhaust cycle.


Time: 45-60 min.
-
Cool S-Gal/LB Agar Blend to approx. 50°C





Add antibiotic supplement.


Note: A 50°C water bath will minimize equilibration time.

Time: 10 min.
Cool agar to approx. 50°C.

Add IPTG solution

Add X-gal to final concentration.

Add antibiotic supplement.


Note: A 50°C water bath will minimize equilibration time.

Time: 10 min.
-
Pour plates.

Note: S-Gal is not light sensitive. No need to protect plates from light.

Time: 10-15 min.
Pour plates.

Note: Plates not used immediately must be protected from light.

Time: 10-15 min.
-
Time saved: 30 min.

For best results, allow plates to dry at room temperature, with lids on, for 16 hours following plating.
(Note: S-Gal/LB Agar Blend can also be prepared using a Microwave Protocol.)

S-Gal is a trademark of Sigma Chemical Co.

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