|Supply Ability:||100 Piece/Pieces per Month|
|Place of Origin:||Jiangsu China|
|Storage:||dry at room temperature (15 ℃ - 25 ℃)|
|name:||Animal Tissue/Cell Genomic DNA Extraction Kit|
|Model Number:||Animal Tissue/Cell Genomic DNA Extraction Kit|
|Packaging Detail:||Plastic bottle: 50T/ bottle Need other packing, please contact customer service|
Animal Tissue/Cell Genomic DNA Extraction Kit
|Place of Origin||Nanjing, China (Mainland)|
|Supply Capacity||100 Piece/per Month|
|Other packaging||please consult customer service|
Before use, please add absolute ethanol into the rinse solution. Please refer to the label on the bottle for adding the rest volume. All centrifugation procedures were performed at room temperature using a desktop centrifuge.
1. Sample treatment:
a. Cells: 1 × 106-1 × 107 cells were collected by centrifugation at 12000 rpm for 1 min. The adherent cells were digested with trypsin, and then were blown into cell suspension by pre cooled PBS. Then the cells were centrifuged at 12000 rpm for 1 min to collect cells. The supernatant was removed as much as possible, and 200 μ l solution a was added to shake until the cells were completely mixed.
b. Tissue: the amount of tissue should not be too large, generally not more than 25mg. Homogenizer can be used for homogenization. It is best to grind it into powder with liquid nitrogen, then fully suspend it with pre cooled PBS or sterile water, and then centrifuge at 12000 rpm for 1 min to collect cells. Remove the supernatant as much as possible, add 200 μ l solution a, and shake until thoroughly mixed.
2. 20 μ l RNase A (10 mg / ml) was added to the suspension and placed at 55 ℃ for 15 min.
3. Adding 20 μ l proteinase K (10mg / ml), fully reversing and mixing, digesting in 55 ℃ water bath, the cell digestion time is shorter, and the tissue digestion time is longer, generally it takes 1-3 hours to complete (rat tail needs to digest overnight). During digestion, the centrifuge tube can be reversed and mixed several times until the sample is completely digested. The indicators of complete digestion are: clear and viscous liquid.
4. Add 200 μ l volume of solution B and mix well. If white precipitate appears, it can be placed at 75 ℃ for 15-30 min. the precipitate will disappear without affecting the follow-up experiment. If the solution is not clear, it means that the sample is not completely digested, which may lead to less and impure DNA extracted, and may also lead to blocking the adsorption column.
5. When 200 μ l of absolute ethanol is added and well mixed, flocculent precipitation may appear, which does not affect DNA extraction. Both the solution and flocculent precipitation can be added to the adsorption column.
6. Centrifuge at 12000 rpm for 1 min, discard the waste liquid, and put the adsorption column into the collection tube.
7. Add 600 μ l rinsing solution to the adsorption column (please check whether absolute ethanol has been added before use), centrifuge at 12000rpm for 1min, discard the waste liquid, and put the adsorption column into the collection pipe.
8. Add 600 μ l rinsing solution into the adsorption column, centrifuge at 12000 rpm for 1 min, discard the waste liquid, and put the adsorption column into the collection pipe.
9. After centrifugation at 12000rpm for 2min, the adsorption column was exposed to room temperature or 50 ℃ for several minutes to remove the residual rinsing solution in the adsorption column. Otherwise, the ethanol in the rinsing solution would affect the subsequent experiments, such as enzyme digestion, PCR, etc.
10. Put the adsorption column into a clean centrifuge tube, add 50-200 μ l of eluent preheated by 65 ℃ water bath into the central suspension of the adsorption membrane, place it at room temperature for 5 min, and centrifuge at 12000 rpm for 2 min.
11. High quality genomic DNA can be obtained by adding the eluent from centrifugation into the adsorption column and centrifuging at 12000 rpm for 2 min.
matters needing attention:
1. After the kit was opened, RNase A and protease K should be stored at - 20 ℃.
2. The sample should avoid repeated freezing and thawing, otherwise the extracted DNA fragment will be smaller and the extraction amount will be decreased.
3. If the solution in the kit precipitates, it can be re dissolved in 65 ℃ water bath before use, without affecting the effect.
4. The volume of eluent buffer should be no less than 50 μ L. if the volume is too small, the recovery efficiency will be affected. The pH value of eluent also affects the elution efficiency. If water is needed as eluent, the pH value should be about 8.0 (NaOH can be used to adjust the pH value of water to this range). If the pH value is lower than 7.0, the elution efficiency will be reduced.
Nanjing Duly Biotech Co.,Ltd
has been established for 11 years. It is mainly engaded in biological and chemical reagents,including enzyme reagents, proteins, antibiotics, plant hormones, nucleic acids, pigments, amino acids, vitamins,carbohydrates, buffers, surfactants,culture media and Separation of materials and kits.There are more than 10,000 products in total.
Our company’s aim is to provide customers with the best quality products and the most intimate service.Personnel has been trained; every batch of material is checked; each procedure guarantees the quality; every bag of our products is top grade;service is thoughtful all the time.The quality, colors and forms of our products have reached a high level in related chemical industry, which relies on characteristic and advanced processing technology and it has gained a high and reliable reputation from customers at both quality and services.