Candida albicans genomic DNA|BNCC356122 |BNCC

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Candida albicans genomic DNA

Culture medium	YM medium (English name: YM): yeast extract 3.0g, malt extract 3.0g, glucose 10.0g, peptone 5.0g, agar 20.0g (without liquid medium), distilled water 1.0L,pH 6.2±0.2. Sterilization at 121 ℃ for 15min.
Subculture procedure	Please add absolute ethanol to buffer GD and rinse solution PW before use. Please refer to the label on the bottle for the added volume; 1. Take yeast cells (no more than 5 × 107cells at most), centrifuge at 12,000rpm(13,400 × g) for 1min, and absorb the supernatant as much as possible (when there is a large amount of bacterial liquid, the bacterial precipitate can be collected into a centrifuge tube through several centrifugations); 2. Breaking of yeast cell wall: Enzymatic method: 600 μL sorbitol buffer is added to thallus, and about 50U Lyticase (customer provided, catalog number: RT410) is added to fully mix. Treatment at 30 ℃ for 30min. Centrifuge at 4000rpm(1500 ×g) for 10min, discard supernatant and collect precipitate. Note: The above is the Lyticase dosage of 5 × 107 yeast cells. According to the different strains of yeast and the number of yeast cells, the concentration of Lyticase used and the incubation time should be adjusted appropriately; 3. Add 200 μL of buffer GA to the precipitate and resuspend the precipitate, and fully mix well. If RNA needs to be removed, 4 μL RNase A(100 mg/ml) solution (customer provided, catalog number: RT405-12) can be added, oscillated for 15sec, and placed at room temperature for 5min; 4. Add 20 μL of Proteinase K solution and mix well; 5. Add 220 μL of buffer GB, mix thoroughly upside down, place at 70 ℃ for 10min, the solution strain is clear, and centrifuge briefly to remove water droplets on the inner wall of the pipe cover. Note: White precipitate may be produced when adding buffer GB, which will disappear when placed at 70 ℃ and will not affect subsequent experiments. If the solution is not clear, it means that the cell lysis is not complete, which may lead to a small amount of extracted DNA and the extracted DNA is not pure; 6. Add 220 μL of anhydrous ethanol and mix it completely upside down. Flocculent precipitation may appear at this time. Brief centrifugation to remove water droplets on the inner wall of the pipe cover; 7. Add the solution and flocculent precipitate obtained in the previous step into an adsorption column CB3 (the adsorption column is put into the collection tube), 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 500 μL of buffer GD to the adsorption column CB3 (please check whether absolute ethanol has been added before use), centrifuge 30sec at 12,000rpm(13,400 ×g), pour out the waste liquid, and put the adsorption column CB3 into the collection tube; 9. Add 600 μL of rinsing solution PW to the adsorption column CB3 (please check whether anhydrous ethanol has been added before use), centrifuge 30sec at 12,000rpm(13,400 ×g), pour out the waste liquid, and put the adsorption column CB3 into the collection pipe; 10. Repeat operation step 9; 11. Put the adsorption column CB3 back into the collection tube, centrifuge at 12,000rpm(13,400 ×g) for 2min, and pour out the waste liquid. The adsorption column CB3 was placed at room temperature for several minutes to thoroughly dry the residual rinse in the adsorption material. Note: The purpose of this step is to remove the residual rinse in the adsorption column. The residual ethanol in the rinse will affect the subsequent enzyme reaction (enzyme digestion, PCR, etc.) experiments; 12. Transfer the adsorption column CB3 into a clean centrifuge tube, add 50-200 μL of elution buffer TE dropwise to the middle part of the adsorption membrane, place it at room temperature for 2-5min, centrifuge at 12,000rpm(13,400 ×g) for 2min, and collect the solution into the centrifuge tube. Note: In order to increase the yield of genomic DNA, the solution obtained by centrifugation can be added to the adsorption column, placed at room temperature for 2min, and centrifuged at 12,000rpm(13,400 ×g) for 2min. The volume of elution buffer should not be less than 50 μL, and too small a volume 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 DNA products should be kept at -20 ℃ to prevent DNA degradation;
Growth conditions	28 ℃, aerobic, 24-48h
Storage conditions	2-8 ℃
application	Research; Analytical testing. The standard strain for the verification and evaluation of the microbial killing effect of disinfection equipment stipulated in the "Disinfection Technical Specification.
Sharing mode	Public welfare sharing

Genomic DNA

Product content: 5μg ;

Purity： (A260/A280):1.7-2.0

Product format: Freeze-dried powder

Storage conditions: 2~8 ℃

Validity period: 6 months

Biosafety level: 1

Purpose: Molecular research

Notes: If you find any abnormal conditions such as damage on the day of receipt, please contact customer service within 24 hours; after opening, it should be used up once and cannot be stored. If it is not opened temporarily, it can be stored at -20°C. Please operate in strict accordance with this instruction, otherwise it will cause abnormality, deactivation, etc., and no reissue service will be provided.

Steps:

1. Prepare sterile ddH₂O;

2. centrifuge the sample tube 12000r /min for 1min, and add the corresponding volume of sterile ddH₂O as required;

3. It can be used normally after a short period of centrifugation after oscillation;

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