sterile talcum powder|BNCC372951 |BNCC

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sterile talcum powder

Subculture procedure	Please add absolute ethanol to the buffer solution GD and rinsing solution PW before use. Please refer to the label on the bottle for the added volume; 1. Take 1-5ml of bacterial culture solution, centrifuge at 10,000rpm(11,500 ×g) for 1min, and suck up the supernatant as much as possible; 2. Add 200 μL buffer GA to the cell pellet and shake until the cell is completely suspended. Note: For gram-positive bacteria that are difficult to break the wall, the second step can be skipped and lysozyme solution can be added for wall breaking treatment. The specific method is to add 110μL buffer (20mM Tris,pH8.0;2mM Na2-EDTA;1.2%Triton) and 70μL lysozyme solution (50 mg/mL, customer-owned, catalog number: RT401) at 37 ℃ for more than 30min. If RNA needs to be removed, 4μL RNase A(100 mg/mL) solution (provided by the customer, catalog number: RT405-12) can be added, shaken for 15sec, and left at room temperature for 5min; 3. Add 20 μL of Proteinase K solution to the tube and mix well; 4. Add 220μL buffer GB, shake for 15sec, place at 70 ℃ for 10min, strain the solution clear, and centrifuge briefly to remove water droplets on the inner wall of the tube cover. Note: White precipitate may be produced when buffer GB is added, which will disappear when placed at 70 ℃, and will not affect subsequent experiments. If the solution does not become clear, it indicates that the cell lysis is not complete, which may lead to a small amount of extracted DNA and impure DNA extracted; 5. Add 220μL anhydrous ethanol, fully shake and mix for 15sec, flocculent precipitation may occur at this time, and centrifuge briefly to remove water droplets on the inner wall of the tube cover; 6. Add the solution and flocculent precipitate obtained in the previous step to an adsorption column CB3 (the adsorption column is placed in the collection tube), centrifuge at 12,000rpm(13,400 ×g) for 30sec, pour out the waste liquid, and place the adsorption column CB3 in the collection tube; 7. Add 500μL buffer GD to the adsorption column CB3 (please check whether anhydrous ethanol has been added before use), centrifuge at 12,000rpm(13,400 ×g) for 30sec, pour out the waste liquid, and put the adsorption column CB3 into the collection tube; 8. Add 600μL of rinsing liquid PW to the adsorption column CB3 (please check whether anhydrous ethanol has been added before use), centrifuge at 12,000rpm(13,400 ×g) for 30sec, pour out the waste liquid, and put the adsorption column CB3 into the collection tube; Repeat step 8; 10. Put the adsorption column CB3 back into the collection tube, centrifuge at 12,000rpm(13,400 x g) for 2min, and pour out the waste liquid. The adsorption column CB3 is left at room temperature for a few minutes to thoroughly dry the residual rinse in the adsorption material. Note: The purpose of this step is to remove the residual rinsing liquid in the adsorption column, and the residual ethanol in the rinsing liquid will affect the subsequent enzyme reaction (enzyme digestion, PCR, etc.) experiment; 11. Transfer the adsorption column CB3 into a clean centrifuge tube, add 50-200μL elution buffer TE to the middle part of the adsorption membrane, place 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 50 μL, which will affect the recovery efficiency. The pH value of the eluent has a great influence on the elution efficiency. If ddH2O is used as eluent, the pH value should be in the range of 7.0-8.5. If the pH value is lower than 7.0, the elution efficiency will be reduced. And the DNA product should be kept at -20 ℃ to prevent DNA degradation. In order to increase the yield of genomic DNA, the solution obtained by centrifugation can be added to the adsorption column CB3, placed at room temperature for 2min, and centrifuged at 12,000rpm(13,400 ×g) for 2min;
Storage conditions	2-8 ℃
Safety level	0
application	Talc fineness:<800 mesh (95% diameter <15 μm). Specific use: Blank control of the experimental method for evaluating the permeability of barrier materials to bacteria-carrying particles.
Sharing mode	Public welfare sharing