Product Description Product Name Generic Name: Nucleic Acid Extraction Kit Commercial Name: Viral DNA/RNA Rapid Extraction Kit (Spin-column) Packing DP4611(50 preps) /DP4612(100 preps) /DP4613(200 preps) Intended Use For isolation and purification of high quality virus DNA/RNA from cells, blood, serum, plasma, lymph, acellular fluid, cell culture supernatant or various virus preserving fluid. Principle The kit applies the unique binding buffer/ Protease K to rapidly lyse nuclease of virus and inactivate virus. Then genome DNA and RNA selectively adsorbs to silicified membrane in high salt solution. Impurities such as cellular metabolite and proteins etc. are removed through a series of of elution- centrifugation steps. Finally purified genome DNA and RNA is eluted from silica membrane by low salt elution buffer EB. Storage and Validity 1. Proteinase K (dry powder) is stored under -20±5ºC; Buffer RW can be stored under 2~8ºC for one month, and -20±5ºC for one year; other components are stored under room temperature. 2. This kit shall be valid for one year. Please use it within its validity. Applicable Instruments This model is suitable for BioTeke CR3180 benchtop high speed refrigerated centrifuge and similar instruments. Requirements for Samples Please use fresh samples or samples that are stored properly. We suggest experiments be performed as soon as the samples are collected. Please store samples at 2~8 ºC temporarily or at -20ºC or -80ºC for long term preservation. Procedures * Please add absolute ethyl alcohol into washing buffer WB as specified before use! Preparation of proteinase K: Add 1000μL of lab water in each piece of proteinase K(dry powder) (the final concentration is 40mg/mL). Mix it upside down ten times until it is completely dissolved. Please store it at 2~8ºC for short-term usage and -20±5ºC for long-term storage. Avoid repeated freeze-thaw for more than five times. 1. Add 200μL of liquid sample(serum, plasma, blood etc.) into a 1.5mL centrifuge tube. For initial volume of liquid sample less than 200μL, please add 1×PBS until it is200μL. If initial volume is between 200μL-300μL, please add reagents proportionally in the subsequent procedures. For tissue samples, please grind first, use 200μL of buffer LB to resuspend, and then add it to virus lysis buffer. 2. Add 500μL of virus lysis buffer VL1. Add 20μL of proteinase K solution (40mg/mL). Shake for 15 seconds for sufficient mixing. Then incubate at 65ºC for 15min. During incubation, please mix it up and down for 3-4 times. 3. Add 300µl absolute ethyl alcohol and then shake it up and down to mix thoroughly. Proper strength and thoroughly mix is very important in the above procedures. Insufficient mixing will result in low yield of nucleic acid. 4. Transfer the solution and flocculated precipitation (if there is) into a spin column VI (spin column in collection tube). Centrifuge at 10,000rpm for 30s, and empty filtrate in collection tube. 5. Add 700µl of buffer RW (please ensure absolute ethyl alcohol is added before use) and centrifuge at 12,000rpm for 30s. Discard filtrate. 6. Add 500µl of buffer RW (please ensure absolute ethyl alcohol is added before use) and centrifuge at 12,000rpm for 30s. Discard filtrate. 7. Put the spin column VI back to the collection tube and centrifuge at 13,000rpm for 2 min. Remove washing buffer if possible. Otherwise the leftover ethanol will affect the next reaction. 8. Transfer the spin column VI to a new centrifuge tube and add 30-50µl preheated (65 ºC -70ºC waterbath ) buffer EB to the middle of adsorption film. Place it at room temperature for 2min. Centrifuge at 12,000rpm for 1min.Add solution into spin column and place it at room temperature for 2min. Centrifuge at 12,000rpm for 1min. 9. DNA can be stored at 2-8ºC, and -20ºC for long-term storage. It is better to reverse transcribe RNA into cfDNA or store RNA at -80ºC. Performance 1. This kit can be used to extract nucleic acid from blood, plasma and serum effectively 2. The results of the kit are stable and have good repeatability. Attention 1. Please add ethanol in buffer RW as specified before use (add 60mL pf ethanol in 15mL RW, or 100mL ethanol in 25mL RW, or 200mL ethanol in 50mL RW), and mix it up thoroughly). Please check to avoid repeated adding!2. To avoid decreased activity and facilitate transportation, protease K is provided as dry powder. After receipt, please centrifuge shortly and add 1mL sterile water to dissolve (the final concentration is 40mg/mL). As repeated freeze and thaw would decrease protease activity. Please split packages (20μL per package) immediately after dissolution and store under -20ºC. 3. Please cover the lid tightly when finished using solutions to avoid volatilization, oxidization and change of PH values after long exposure to air.