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Cloning & Expression
S-Gal Preparation
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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. |
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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. |
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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. |
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| 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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