Candida albicans Genomic DNA - Basic Information

Culture Medium	Special peptone: 23.0g, soluble starch: 1.0g, sodium chloride: 5.0g, agar: 10.0g, defibrinated sheep blood: 5% (added at 50°C), pH: 7.3±0.2 (25°C)
Growth Conditions	37°C; 48-72h; Microaerophilic
Storage Conditions	2-8°C
Procedure	Before use, add anhydrous ethanol to Buffer GD and Wash Solution PW, referring to the bottle labels for the required volume. 1. Take yeast cells (no more than 5x10^7 cells), centrifuge at 12,000rpm (13,400xg) for 1min, and carefully remove the supernatant (for larger volumes, centrifuge multiple times to collect the cell pellet in a single tube). 2. Disruption of yeast cell walls: Enzymatic method: Add 600μL sorbitol buffer to the cell pellet, add approximately 50U Lyticase (custom-supplied, catalog number: RT410), mix thoroughly. Incubate at 30°C for 30min. Centrifuge at 4,000rpm (1,500xg) for 10min, discard the supernatant, and collect the pellet. Note: The above is the Lyticase amount for 5x10^7 yeast cells; adjust the Lyticase concentration and incubation time according to the yeast strain and cell count. 3. Add 200μL Buffer GA to the pellet, resuspend, and mix thoroughly. If RNA removal is needed, add 4μL RNase A (100mg/mL) solution (custom-supplied, catalog number: RT405-12), vortex for 15sec, and let stand at room temperature for 5min. 4. Add 20μL Proteinase K solution and mix. 5. Add 220μL Buffer GB, mix thoroughly by inversion, incubate at 70°C for 10min until the solution becomes clear, and briefly centrifuge to remove droplets from the tube cap. Note: A white precipitate may form when adding Buffer GB, which usually disappears at 70°C and will not affect subsequent experiments. If the solution does not become clear, cell lysis may be incomplete, potentially resulting in low DNA yield and impurity. 6. Add 220μL anhydrous ethanol, mix thoroughly by inversion (flocculent precipitate may form), and briefly centrifuge to remove droplets from the tube cap. 7. Transfer the solution and precipitate from the previous step into a CB3 adsorption column (placed in a collection tube), centrifuge at 12,000rpm (13,400xg) for 30sec, discard the waste liquid, and place the CB3 column back into the collection tube. 8. Add 500μL Buffer GD to the CB3 column (ensure anhydrous ethanol has been added before use), centrifuge at 12,000rpm (13,400xg) for 30sec, discard the waste liquid, and place the CB3 column back into the collection tube. 9. Add 600μL Wash Solution PW to the CB3 column (ensure anhydrous ethanol has been added before use), centrifuge at 12,000rpm (13,400xg) for 30sec, discard the waste liquid, and place the CB3 column back into the collection tube. 10. Repeat step 9. 11. Place the CB3 column back into the collection tube, centrifuge at 12,000rpm (13,400xg) for 2min, discard the waste liquid. Let the CB3 column stand at room temperature for several minutes to thoroughly dry any residual wash solution in the adsorption material. Note: This step aims to remove residual wash solution from the column, as residual ethanol in the wash solution can affect subsequent enzymatic reactions (e.g., digestion, PCR). 12. Transfer the CB3 column into a clean centrifuge tube, add 50-200μL Elution Buffer TE dropwise to the center of the adsorption membrane, let stand at room temperature for 2-5min, centrifuge at 12,000rpm (13,400xg) for 2min, and collect the solution into the centrifuge tube. Note: To increase genomic DNA yield, the collected solution can be added back onto the column, let stand at room temperature for 2min, and centrifuged at 12,000rpm (13,400xg) for 2min. The elution buffer volume should not be less than 50μL, as smaller volumes reduce recovery efficiency. The pH of the elution buffer significantly affects elution efficiency. If using ddH2O as the elution buffer, ensure the pH is between 7.0-8.5; a pH below 7.0 will reduce elution efficiency. DNA products should be stored at -20°C to prevent degradation.
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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