Staphylococcus aureus Genomic DNA - Basic Information

Culture Medium	Peptone: 10.0g, Beef powder: 3.0g, Sodium chloride: 5.0g, Agar: 15.0g, pH value: 7.3±0.1 (25°C)
Growth Conditions	37°C, aerobic, 18-24h.
Growth Characteristics	Gram-positive cocci, arranged singly, in pairs, or in irregular clusters. No spore formation. Ornithine decarboxylase negative, ferments glucose, maltose, lactose, and sucrose to produce acid without gas, ferments mannitol. Methyl red reaction positive, Voges-Proskauer test weakly positive. Transparent hemolytic rings form around colonies on blood agar plates. Produces golden pigment. Facultative anaerobic, optimum pH 7.0-7.5.
Storage Conditions	2-8°C
Morphology	Round, with neat edges, opaque, light in color, convex in the center, smooth and shiny surface, moist texture, easy to pick up, no pigment production, Gram-positive (blue-purple), cocci, purity: pure
Procedure	Add anhydrous ethanol to buffer GD and rinse solution PW before use, with the volume as indicated on the bottle labels. 1. Take 1-5ml of bacterial culture solution, centrifuge at 10,000rpm (11,500×g) for 1min, and remove the supernatant as completely as possible. 2. Add 200μL of buffer GA to the bacterial pellet and vortex until the bacteria are fully suspended. Note: For Gram-positive bacteria with tough cell walls, skip step 2 and use lysozyme solution for wall digestion. The specific method is: add 110μL buffer (20mM Tris, pH8.0; 2mM Na2-EDTA; 1.2% Triton) and 70μL lysozyme solution (50mg/mL, prepared by the user, Cat. No.: RT401), and incubate at 37°C for more than 30min. If RNA removal is needed, add 4μL RNase A (100mg/mL) solution (prepared by the user, Cat. No.: RT405-12), vortex for 15sec, and let stand at room temperature for 5min. 3. Add 20μL Proteinase K solution to the tube and mix well. 4. Add 220μL buffer GB, vortex for 15sec, and incubate at 70°C for 10min until the solution becomes clear. Briefly centrifuge to remove droplets from the inner wall of the tube cap. Note: A white precipitate may form when buffer GB is added, which usually disappears at 70°C and does not affect subsequent experiments. If the solution does not become clear, cell lysis may be incomplete, leading to low DNA yield and impure DNA extraction. 5. Add 220μL anhydrous ethanol, vortex thoroughly for 15sec. A flocculent precipitate may appear at this point. Briefly centrifuge to remove droplets from the inner wall of the tube cap. 6. Transfer the solution and flocculent precipitate obtained in the previous step into an adsorption column CB3 (placed in a collection tube), centrifuge at 12,000rpm (13,400×g) for 30sec, discard the waste liquid, and place the adsorption column CB3 back into the collection tube. 7. Add 500μL buffer GD to the adsorption column CB3 (check if anhydrous ethanol has been added before use), centrifuge at 12,000rpm (13,400×g) for 30sec, discard the waste liquid, and place the adsorption column CB3 back into the collection tube. 8. Add 600μL rinse solution PW to the adsorption column CB3 (check if anhydrous ethanol has been added before use), centrifuge at 12,000rpm (13,400×g) for 30sec, discard the waste liquid, and place the adsorption column CB3 back into the collection tube. 9. Repeat step 8. 10. Place the adsorption column CB3 back into the collection tube, centrifuge at 12,000rpm (13,400×g) for 2min, and discard the waste liquid. Leave the adsorption column CB3 at room temperature for several minutes to thoroughly air-dry any residual rinse solution in the adsorption material. Note: This step aims to remove residual rinse solution from the adsorption column, as ethanol residue in the rinse solution can affect subsequent enzymatic reactions (e.g., enzyme digestion, PCR). 11. Transfer the adsorption column CB3 into a clean centrifuge tube, add 50-200μL elution buffer TE dropwise onto the center of the adsorption membrane, let stand at room temperature for 2-5min, centrifuge at 12,000rpm (13,400×g) for 2min, and collect the solution into the centrifuge tube. Note: The volume of elution buffer should not be less than 50μL, as too small a volume reduces recovery efficiency. The pH of the elution buffer significantly affects elution efficiency. If ddH2O is used as the eluent, ensure its pH is within the range of 7.0-8.5; a pH below 7.0 will reduce elution efficiency. DNA products should be stored at -20°C to prevent degradation. To increase the yield of genomic DNA, the collected solution can be reapplied to the adsorption column CB3, let stand at room temperature for 2min, and centrifuged at 12,000rpm (13,400×g) for 2min.
Reference Applications	Research; analytical testing. Assay of antibiotics, test microorganism for medical device sterilization efficacy testing as per the "Technical Standard for Disinfection", positive control strain for "GB 4789.2-2010 Enumeration of Total Bacterial Count", quality control strain in the 2015 edition of the Chinese Pharmacopoeia. Sterility testing; determination of neomycin, cefactor, amikacin, kanamycin, cefoxitin, cefazolin, nafcillin, vancomycin, lincomycin, penicillin, tobramycin, neomycin B, spiramycin, gramicidin, cefotaxime, cephalexin, cephalosporin, mithramycin, gentamicin, doxycycline, methicillin, cefamandole, cephalosporin, cloxacillin, cycloserine, erythromycin, tetracycline, methacycline, cephalosporin, dicloxacillin, demeclocycline, oxytetracycline, oxytetracycline rolitetracycline, and chlortetracycline; drug sensitivity tests for kanamycin, clindamycin, streptomycin, and tetracycline. GB 4789.2-2016 Enumeration of Total Bacterial Count.
Sharing Method	Public welfare sharing

📋 Notes:

1. Storage Conditions:Lyophilized powder should be stored at 2–8°C away from light. After reconstitution or for solution-type products, store at –20°C and avoid repeated freeze-thaw cycles. For short-term frequent use (within 1 week), temporary storage at 4°C is acceptable.
2. Reconstitution Method:Prior to use, briefly centrifuge the lyophilized powder (e.g., 12,000 rpm for 1 minute) to collect the DNA at the bottom of the tube. It is recommended to dissolve in sterile TE buffer (pH 8.0) or sterile ddH₂O (pH 7.0–8.5). Mix gently; avoid vigorous vortexing.
3. Contamination Prevention:Gloves must be worn during operation. Use sterile, nuclease-free consumables. It is recommended to aliquot the reconstituted DNA into single-use portions before freezing, to prevent degradation caused by repeated freeze-thaw cycles and to avoid nuclease contamination.
4. Experimental Suitability:This product is suitable for common molecular biology applications such as PCR, qPCR, restriction enzyme digestion, and library construction. If the downstream assay is sensitive to inhibitors, ensure that the DNA solution is free of residual ethanol or high-concentration salt interference.
5. Intended Use Statement:The product is for research use only and is not intended for clinical diagnostic purposes.

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