Dextrose Starch Agar | Medium for Amylolytic Microorganisms | AS‑1204
Dextrose Starch Agar (DSA)
GC Agar Base & General-Purpose Nutrient Medium | Neisseria, Starch Hydrolysis, Heterotrophic Cultivation | Cat. No. AS-1204
Dextrose Starch Agar (DSA, AS-1204) is a versatile nutrient medium used primarily as a GC Agar Base for the cultivation of Neisseria gonorrhoeae and N. meningitidis when supplemented with haemoglobin and growth factors (IsoVitaleX or equivalent). It is also widely used for starch hydrolysis testing (amylase activity detection with iodine flooding), general heterotrophic bacterial cultivation, and colony morphology differentiation in environmental, food, and clinical microbiology laboratories.
The unique formula combines a rich proteose peptone nitrogen source with soluble starch as both a hydrolysis substrate and energy reserve, gelatin as an additional protein source, disodium phosphate as a pH buffer, and low dextrose (2 g/L) for carbohydrate supplementation without the intense fermentation seen in high-sugar media.
2. Starch Hydrolysis Testing (differential use): Soluble starch (10 g/L) is the substrate for amylase-producing bacteria. After incubation, flood the plate with Gram's iodine solution. Starch-positive organisms (amylase producers) show a clear halo around the colony where starch has been hydrolysed. Starch-negative organisms remain surrounded by the blue-black iodine-starch complex. This is used for identification of Bacillus, Clostridium, Pseudomonas, and environmental isolates.
3. General Nutrient Base: Without supplementation, DSA supports broad heterotrophic bacterial growth from food, water, soil, and industrial samples. The gelatin (20 g/L) also allows gelatinase testing — pour melted agar into gelatin agar medium or observe liquefaction.
Positive: Clear/white halo around colony = starch hydrolysed by amylase.
Negative: Blue-black colouration around and under colony = starch intact.
1. Prepare DSA base at 65 g/L. Autoclave 121°C / 15 min. Cool to 50–55°C.
2. Add aseptically: 2% haemoglobin solution (equal volume, pre-sterilised) to form chocolate agar colour.
3. Add aseptically: IsoVitaleX Enrichment (or Supplement B) at 1% v/v — supplies NAD, glutamine, coenzymes, thiamine pyrophosphate.
4. Mix gently and pour plates immediately. Incubate inoculated plates at 35–37°C in 5% CO₂ for 24–48h.
Result: N. gonorrhoeae grows as small, translucent, glistening colonies on the chocolate-coloured medium.
| Medium | Key Components | Starch | Gelatin | Best For |
|---|---|---|---|---|
| DSA (AS-1204) ★ | Proteose Peptone 15g + Starch 10g + Gelatin 20g | ✓ 10 g/L | ✓ 20 g/L | GC Agar Base; starch hydrolysis; heterotrophs |
| Nutrient Agar (AS-1309) | Peptone 5g + Beef Extract 3g | ✕ | ✕ | Non-fastidious general cultivation |
| Tryptic Soy Agar (AS-1371) | Casein 15g + Soy 5g + NaCl 5g | ✕ | ✕ | USP/EP sterility, general enrichment |
| Starch Agar (plain) | Peptone + Starch only | ✓ | ✕ | Starch hydrolysis only; no gelatin |
| Plate Count Agar (AS-1329) | Tryptone 5g + Yeast Extract 2.5g + Glucose 1g | ✕ | ✕ | Total viable count (TVC) — food/water |
| Manufacturer | Product Name | Cat. No. |
|---|---|---|
| Sigma-Aldrich / Merck | Dextrose Starch Agar (GC Medium Base) | D3060 |
| Merck Millipore | GC Agar Base | 1.10210 |
| HiMedia | Dextrose Starch Agar | M263 |
| Thermo Scientific / Oxoid | GC Agar Base | CM0367 |
| Product Name | Dextrose Starch Agar (DSA) |
| Catalogue Number | AS-1204 |
| Synonyms | GC Agar Base; GC Medium Base; Chocolate Agar Base; Dextrose Starch Medium; Neisseria Culture Agar Base |
| Commercial Equivalents | Sigma-Aldrich D3060 | Merck 1.10210 | HiMedia M263 | Oxoid CM0367 |
| Medium Type | General-purpose nutrient agar; GC Agar Base (when supplemented) |
| Dissolution | 65 g/L in distilled or deionised water |
| Final pH at 25°C | 7.2 ± 0.2 |
| Sterilisation | Autoclave 121°C for 15 minutes |
| Appearance (powder) | Light beige to cream, free-flowing homogeneous powder |
| Appearance (prepared agar) | Pale amber, clear to slightly opalescent; firm gel |
| Storage (powder) | 15–30°C, dry, tightly sealed, away from direct sunlight |
| Storage (prepared plates) | 2–8°C, inverted, protected from light; use within 2 weeks |
| HS Tariff Code | 3821.00.00 |
| Ingredient | g / L | Function |
|---|---|---|
| Proteose Peptone | 15.0 | Primary nitrogen source — long-chain peptides, amino acids, growth factors for fastidious organisms |
| Soluble Starch | 10.0 | Polysaccharide energy reserve; substrate for amylase (starch hydrolysis) testing |
| Gelatin | 20.0 | Additional protein/nitrogen source; substrate for gelatinase activity testing |
| Sodium Chloride (NaCl) | 5.0 | Osmotic balance — physiological ionic strength |
| Disodium Phosphate (Na₂HPO₄) | 3.0 | pH buffer — maintains pH 7.2 ± 0.2 during growth |
| Dextrose (D-Glucose) | 2.0 | Supplementary carbon/energy source; low level avoids excessive acid production |
| Agar | 10.0 | Solidifying agent (lower than standard 15 g/L — softer gel for colonial spread) |
| Total | 65.0 g/L | pH 7.2 ± 0.2 at 25°C — Autoclave 121°C / 15 min |
| Organism (ATCC) | Application | Incubation | Expected Result |
|---|---|---|---|
| Neisseria gonorrhoeae (49226) | GC Agar Base (+ supplements) | 35°C / 5% CO₂ / 24–48h | Small, translucent, glistening colonies ✓ |
| Escherichia coli (25922) | General growth (plain DSA) | 35°C / 18–24h | Good growth; starch hydrolysis negative ✓ |
| Bacillus subtilis (6633) | Starch hydrolysis test | 35°C / 24–48h | Good growth; clear halo with iodine (positive) ✓ |
| Staphylococcus aureus (25923) | General growth (plain DSA) | 35°C / 18–24h | Good growth; starch hydrolysis negative ✓ |
| # | Reference |
|---|---|
| 1 | Sigma-Aldrich / Merck. Dextrose Starch Agar (GC Medium Base), Cat. D3060. Specification sheet; current edition. [Reference formula] |
| 2 | Murray PR, et al. Manual of Clinical Microbiology. 12th ed. ASM Press; 2019. [GC Agar for Neisseria culture] |
| 3 | CLSI. Performance Standards for Antimicrobial Susceptibility Testing of Neisseria gonorrhoeae. CLSI M100; current edition. |
| 4 | Atlas RM. Handbook of Microbiological Media. 4th ed. CRC Press; 2010. [Dextrose Starch Agar and starch hydrolysis methodology] |
| 5 | ISO 11133:2014. Culture media performance testing. Geneva: ISO; 2014. |
| Product Name | Dextrose Starch Agar (DSA) |
| Catalogue No. | AS-1204 |
| Supplier | AuSaMicS Pty Ltd | ABN 56 676 640 467 |
| Address | 31 Longview CT, Thomastown VIC 3074, Australia |
| Emergency | Poisons Information Centre: 13 11 26 (24 hr) |
| Phone | +61 412 520 598 | support@ausamics.com.au |
| GHS Classification | NOT classified as a hazardous substance under Australian WHS Regulations 2023 at intended use concentrations. |
| Signal Word | None required |
| Other Hazards | Combustible dry powder. Dust may cause mild respiratory irritation. No significant chemical hazard at normal laboratory handling levels. |
| Component | g/L | CAS | Hazard |
|---|---|---|---|
| Proteose Peptone | 15.0 | 73049-73-7 | Not hazardous |
| Soluble Starch | 10.0 | 9005-84-9 | Not hazardous |
| Gelatin | 20.0 | 9000-70-8 | Not hazardous |
| Sodium Chloride | 5.0 | 7647-14-5 | Not hazardous |
| Disodium Phosphate | 3.0 | 7558-79-4 | Not hazardous |
| Dextrose | 2.0 | 50-99-7 | Not hazardous |
| Agar | 10.0 | 9002-18-0 | Not hazardous |
| Respiratory PPE | P1 filter when weighing bulk powder |
| Eye Protection | Safety glasses when handling dry powder |
| Skin / Gloves | Nitrile gloves when handling inoculated cultures |
| Waste (plain DSA) | Autoclave inoculated plates (121°C / 15 min) before disposal as general microbiological waste |
| Transport | Not dangerous goods — ADG, IMDG, IATA |
| Regulatory | Australian WHS Regulations 2023 | GHS 7th Edition | AICIS compliant |
| Parameter | Specification | Method |
|---|---|---|
| Appearance (powder) | Light beige to cream, free-flowing, homogeneous | Visual |
| Appearance (prepared agar) | Pale amber, clear to slightly opalescent; firm gel | Visual after dissolution |
| pH (prepared, 25°C) | 7.2 ± 0.2 | pH meter (calibrated) |
| Dissolution | Complete at 65 g/L with boiling | Visual |
| Moisture Content | ≤5.0% (w/w) | Loss on drying |
| Growth — E. coli ATCC 25922 | Good growth at 35°C / 18–24h; ≤100 CFU inoculum | ISO 11133:2014 |
| Growth — S. aureus ATCC 25923 | Good growth at 35°C / 18–24h; ≤100 CFU inoculum | ISO 11133:2014 |
| Starch hydrolysis function | Clear halo with iodine around B. subtilis ATCC 6633 colonies | Iodine flooding test |
| Batch COA | Available every production lot | Included with order |
31 Longview CT, Thomastown VIC 3074, Australia | ABN 56 676 640 467
✓ Formulated per Sigma-Aldrich D3060 / Merck 1.10210 reference specification
✓ Batch-level QC per ISO 11133:2014 — pH, growth promotion, appearance
✓ COA, TDS, SDS included with every order
✓ Australian stock — same-week dispatch, no import delays
✓ Technical support from our microbiology team