Oil DNA Extraction Kit (10 times, 10ml each time) Instruction Manual
Oil DNA Extraction Kit (10 times, 10ml each time)
Brief Description: This kit is capable of extracting trace amounts of DNA from refined oil for various testing purposes.
Scope of application: Various refined oil extraction time: 40 minutes to one hour, it is the shortest kit in China.
DNA quality: Resin extraction, which can extract high quality DNA from large or micro volume oils, because during the process of refining oil, DNA
The damage is serious, so the DNA is not a single sputum DNA fragment, and the DNA fragment size is distributed in the tens of bp to
Between 600bp, after 1.2% Agarose electrophoresis, DNA may not be visible under UV light, even if it is visible.
Kit composition:
1. Solution A (oil removal buffer): 110ml 2. Solution B (lysis buffer): 120ml
3. Solution C (resin, resin): 550ul (shake well before use)
4. Solution D (washing buffer): 110ml 5. Solution E (TE buffer): 1.5ml
Operation method:
method one:
1. Take 10ml refined oil plus 10 ml solution A, 5ml solution B, shake well, let stand, carefully remove the upper layer
2. Add 50ul of solution C (shake well before use), shake gently and let stand for 2 minutes.
3. Centrifuge at 8000 rpm for 5 minutes (normal temperature or 4 degrees), carefully discard the supernatant.
4. Add 1 ml of solution B, mix well, transfer to a 1.5 ml centrifuge tube, centrifuge at 8000 rpm for 2 minutes, and discard the sputum.
5. Add 1 ml of solution D, mix well, transfer to a 1.5 ml centrifuge tube, centrifuge at 8000 rpm for 2 minutes, and discard the sputum.
6. Repeat step 5
7. Centrifuge at 8000 rpm for 1 minute, blot the supernatant, and allow to dry at room temperature for about 5-10 minutes.
8. Add solution E 50-100ul, mix, 50-60 degree water bath for 2-5 minutes,
9. Centrifuge at 8000-12000 rpm for 2 minutes and take the supernatant as DNA.
Method Two:
1 Take 3ml refined oil plus 3 ml solution A, 3ml solution B, shake well, let stand, carefully remove the upper layer
2 Add 30 ul of solution C (shake well before use), shake gently and let stand for 2 minutes.
Centrifuge at 3 8000 rpm for 5 minutes (normal temperature or 4 degrees), carefully discard the supernatant,
4 Add 3OOul of solution B, mix well, transfer to a 1.5ml centrifuge tube, centrifuge at 8000rpm for 2 minutes, and discard the sputum.
5 Add 500 ul of solution D, mix well, transfer to a 1.5 ml centrifuge tube, centrifuge at 8000 rpm for 2 minutes, and discard the sputum.
6 Repeat step 5
7 8000 rpm, centrifuge for another 1 minute, blot the supernatant, and let it dry for about 5-10 minutes at room temperature.
8 Add solution E 30-50ul, mix, 50-60 degree water bath for 2-5 minutes,
Centrifuge for 2 minutes at 9 8000-12000 rpm and take the supernatant as DNA.
Note: The DNA quality can be detected by PCR. The DNA extracted by Method 1 or Method 2 only needs 3-8ul/reaction.
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