BCG Mycobacterium bovis Genomic DNA (Qubit Measured) - Basic Information

Storage Conditions	2-8°C
Procedure	Before use, add anhydrous ethanol to Buffer GD and Wash Buffer PW; refer to the label on the bottle for the amount. 1. Take 1-5 ml of bacterial culture, centrifuge at 10,000 rpm (11,500×g) for 1 min, and remove the supernatant as much as possible. 2. Add 200 μL Buffer GA to the bacterial pellet and vortex until completely resuspended. Note: For Gram-positive bacteria with tough cell walls, skip step 2 and add lysozyme solution for cell wall digestion as follows: Add 110 μL buffer (20 mM Tris, pH 8.0; 2 mM Na2-EDTA; 1.2% Triton) and 70 μL lysozyme solution (50 mg/mL, provided by user, Cat. No.: RT401), incubate at 37°C for over 30 min. If RNA removal is needed, add 4 μL RNase A (100 mg/mL) solution (provided by user, Cat. No.: RT405-12), vortex for 15 sec, and leave at room temperature for 5 min. 3. Add 20 μL Proteinase K solution to the tube and mix well. 4. Add 220 μL Buffer GB, vortex for 15 sec, incubate at 70°C for 10 min; the solution should become clear. Briefly centrifuge to remove droplets from the inside of the cap. Note: Adding Buffer GB may produce a white precipitate, which usually disappears at 70°C and will not affect subsequent experiments. If the solution does not become clear, cell lysis is incomplete, which may result in low DNA yield and impure extracted DNA. 5. Add 220 μL anhydrous ethanol, vortex thoroughly for 15 sec; at this point, a flocculent precipitate may appear. Briefly centrifuge to remove droplets from the inside of the cap. 6. Transfer the solution and flocculent precipitate from the previous step into a spin column CB3 (placed in a collection tube), centrifuge at 12,000 rpm (13,400×g) for 30 sec, discard the waste liquid, and place the spin column CB3 back into the collection tube. 7. Add 500 μL Buffer GD to the spin column CB3 (ensure anhydrous ethanol has been added beforehand), centrifuge at 12,000 rpm (13,400×g) for 30 sec, discard the waste liquid, and place the spin column CB3 back into the collection tube. 8. Add 600 μL Wash Buffer PW to the spin column CB3 (ensure anhydrous ethanol has been added beforehand), centrifuge at 12,000 rpm (13,400×g) for 30 sec, discard the waste liquid, and place the spin column CB3 back into the collection tube. 9. Repeat step 8. 10. Place the spin column CB3 back into the collection tube, centrifuge at 12,000 rpm (13,400×g) for 2 min, discard the waste liquid. Leave the spin column CB3 at room temperature for a few minutes to thoroughly dry any residual wash buffer from the adsorbent material. Note: This step aims to remove residual wash buffer from the spin column, as ethanol residue in the wash buffer can affect subsequent enzymatic reactions (such as enzyme digestion or PCR). 11. Transfer the spin column CB3 into a clean centrifuge tube, carefully add 50-200 μL Elution Buffer TE to the center of the membrane, leave at room temperature for 2-5 min, centrifuge at 12,000 rpm (13,400×g) for 2 min, and collect the solution in the centrifuge tube. Note: The elution buffer volume should not be less than 50 μL; too small a volume reduces recovery efficiency. The pH of the elution buffer greatly affects elution efficiency. If ddH2O is used as the elution buffer, ensure its pH is within the range of 7.0-8.5; a pH below 7.0 reduces elution efficiency. DNA products should be stored at -20°C to prevent degradation. To increase genomic DNA yield, the collected solution can be reapplied to the spin column CB3, left at room temperature for 2 min, and centrifuged at 12,000 rpm (13,400×g) for 2 min.
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.