Face mask machine Jiangsu Health Medical Technology Development Co., Ltd. , https://www.medicalhos.com
Kit composition:
1. Solution A 25ml 5×STMT (diluted with sterile double distilled water at a volume ratio of 1:4)
2. Solution B 17ml
3. Solution C 0.6ml RNase A
4. Solution D 44ml
5. Solution E 1.25ml Resin (mix well when used)
6. Solution F 30ml lotion (add 40ml of absolute alcohol when used, and mark it in the mouth of the cap)
7. Solution G 5ml TE buffer experimental steps:
(a) fresh anticoagulation or frozen anticoagulation
1. Take 300 μl of whole blood, add 900 μl of solution A, and mix gently 5 to 6 times.
2. Centrifuge at room temperature or ice for 10 min, ≥ 8000 rpm for 20 to 30 seconds, discard the supernatant.
Note: 1) If the red blood cells are not completely dissolved, the precipitate will be red, and 900 μl of solution A may be added, and the above steps are repeated once.
2) If whole blood is frozen before use, repeat steps 1 and 2 twice.
3. Add 300 μl of solution B, gently shake or repeatedly pipette the white precipitate 4 to 6 times.
Add 10 μl of solution C and let stand for 10 min at room temperature.
4. Add 800 μl of solution D, 20 μl of solution E, mix and let stand for 5 min at room temperature.
5. Centrifuge for 30 s at 8000 rpm and discard the supernatant.
6. Add 500 μl of solution F, mix well, centrifuge at ≥ 8000 rpm for 30 to 60 s, and discard the supernatant.
7. Repeat step 6.
8. Centrifuge for 1 min at ≥12,000 rpm and blot the remaining liquid. Leave at room temperature for 5 to 10 minutes.
9. Add 50 ~ 100μl solution G, mix well, heat at 50 ° C for 5 ~ 10min, ≥ 12000rpm centrifuge for 1min, absorb the supernatant,
It is DNA.
(2) Separated white blood cells
1. Take 100 μl of white blood cells, add solution B directly at a ratio of 1:3, mix well, add 10 μl of solution C, and let stand for 10 min at room temperature.
2. The rest is connected to step 4 of (a).
Note:
1. The user can adjust the amount of solution G as appropriate according to the concentration of DNA.
2. The quality of DNA extracted by this method is very high. It is suitable for any experimental requirement of molecular biology. OD260/280≥1.5, the general condition can reach 1.7--1.8. If the ratio is low, it may be hemolysis or storage time is too long.
Whole Blood Genomic DNA Extraction Kit (50 times) - Instructions
Whole blood genomic DNA extraction kit (50 times)