Aquatic Animal Disease Genomic DNA/RNA Extraction Kit I. Introduction The aquatic animal disease genomic DNA/RNA extraction kit is suitable for rapidly extracting high-purity disease genomic nucleic acids from aquatic animal tissue homogenates. The kit is based on silica gel column purification technology. It does not require the use of toxic phenol chloroform extraction or time-consuming alcohol precipitation. The obtained nucleic acid can be directly used in PCR/RT-PCR, Sorter hybridization/Northern hybridization, and Downstream experiments in series such as LAMP/RT-LAMP. Component Content Tube A 48 tubes B 48 lysates 30ml 4. Shelf Life Aquatic Animal Disease Genomic DNA/RNA extraction kit components can be stored dry for 12 months at room temperature (15-25 ° C). It should be placed at 2-8 °C for long-term storage. 5. Materials and tools to be prepared ï¬ Sterile physiological saline ï¬ Anhydrous ethanol ï¬ Clean tweezers and scissors ï¬ Homogenizer, homogenized bag or disposable grinding rod ï¬ Micropipette (100-1000μl, 10-100μl) VII. Frequently Asked Questions This list may help you solve the problems encountered during the extraction process. If you have questions or have any suggestions for the kit, or if you have problems with molecular biology experiments, please contact us. We will do our best to help you solve problems. Symptom Cause Solution Low or no nucleic acid yield The sample is repeatedly thawed to avoid repeated freezing and thawing of the sample. It is recommended to use fresh samples or samples that have only been thawed once. Disposable Surgical Gown,Medical Isolation Gown,Hospital Disposable Surgical Gown,Nonwoven Disposable Surgical Gown Xinxiang Huaxi Sanitary Materials Co., Ltd. , https://www.huaximedical.com
Second, the principle The silica gel column used in this kit is based on a high-strength glass fiber filter. The filter membrane can adsorb nucleic acids by physicochemical interactions such as hydrogen bonding and static electricity under the condition of high concentration ionizing agent (such as guanidine hydrochloride or guanidinium isothiocyanate), while proteins and other impurities are not adsorbed and removed. The nucleic acid-adsorbing filter is washed to remove residual proteins and salts, and finally the DEPC can be used to treat the nucleic acid adsorbed on the water-eluting filter. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.
The aquatic animal disease genomic DNA/RNA extraction kit is based on a silica gel column purification method. The sample is cleaved in the lysate and the nucleic acid is released into the lysate. The nucleic acid is adsorbed on the membrane of the silica gel column through a silica gel column in a high salt environment, and the protein is not adsorbed and removed. The salt is then washed by washing to remove the salt, and finally the nucleic acid is eluted by the nuclease-free eluate.
Third, the composition 071011M
Washing liquid 10ml
Eluent 36ml
Instructions 1 Note: The washing solution is added to 40 ml of absolute ethanol before the initial use and stored at room temperature.
ï¬ Sterilized centrifuge tubes without DNase and RNase (1.5ml or 2ml)
ï¬ Sterilized Tip head without DNase and RNase ï¬ vortex oscillator ï¬ centrifuge (speed ≥ 10,000 rpm)
6. Rapid extraction of DNA/RNA from aquatic animal diseases This protocol uses centrifugation and is suitable for rapid extraction of disease genomic DNA/RNA from aquatic animal tissue samples. The following steps are all performed at room temperature.
1. Take appropriate amount of fresh sample tissue to be tested and add 1~5 times the volume of normal saline for homogenization.
2. Take 200 μl of the homogenate into a 1.5 ml centrifuge tube, add 500 μl of the lysate to the supernatant, mix by inversion, let stand for 5 min to lyse the disease, and centrifuge for 3 min.
3. Tube A was placed in 2 ml tube B, 400 μl was pipetted into tube A, 200 μl of absolute ethanol was added to tube A, and vortexed for 20 seconds.
Centrifuge at 10,000 rpm for 1 min.
5. The filtrate in tube B was discarded, tube A was returned to tube B, 600 μl of washing solution was added to tube A, and centrifuged at 10,000 rpm for 1 min.
6. The filtrate in tube B was discarded, tube A was returned to tube B, and centrifuged at 10,000 rpm for 2 min.
7. Tube A was placed in a new 1.5 ml centrifuge tube. 50 μl of the eluate was added to the tube A, allowed to stand for 1-2 min, and centrifuged at 10,000 rpm for 1 min, and the resulting solution was a disease genomic nucleic acid solution. If not used temporarily, store at -80 °C.
The disease titer in the sample is too low. Use a small concentration column to concentrate the disease sample or increase the amount of the sample.
Insufficient sample homogenization or insufficient lysis. Thoroughly homogenize the sample and prolong the lysis time appropriately.
The washing solution is not diluted with ethanol. Before the initial use, the washing liquid must be added with absolute ethanol to dilute the nucleic acid. The downstream application result is not ideal. The nucleic acid concentration is too low. The amount of DEPC water at the time of elution is reduced to increase the concentration of nucleic acid.
Nucleic acid yield is too low or not See above Ethanol contamination Make sure to follow the conditions in the instructions, such as 10,000 rpm for empty column centrifugation and 1 min for centrifugation.
If the above solution still does not solve your problem, please contact us.