5 min Swab/Saliva DNA Extraction Kit
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- Douglas Oswald French
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1 5 min Swab/Saliva DNA Extraction Kit Catalog #; D-1007 The fastest and simplest but most reliable Nucleic Acid extraction system Purification of total DNA from buccal swab, saliva, cervical swab, milk, and semen Proteinase K treatment is required for better quality of DNA. From proteinase K treated sample to DNA extraction in 5 minutes Single column format Final 2-15 ug of genomic DNA The kit can be stored at ambient temperature for more than a year. One kit for 50 DNA purification The extracted DNA will show around ratio of 260/280. The extracted DNA can be used in any downstream steps without any inhibitory effect. 5 min Swab/Saliva DNA Extraction Kit
2 Storage Conditions and Product Stability All components should be kept tightly sealed and stored at room temperature. These reagents should remain stable for at least 1 year in their unopened containers. Precautions and Disclaimers This kit is designed for research purposes only. It is not intended for human or diagnostic use. Customer-Supplied Reagents and Equipment Benchtop microcentrifuge 1.5 ml microcentrifuge tubes Vortex % ethanol % isopropanol Proteinase K (2 mg/ml) Heatblock set at 65 o C Proteinase K (20 mg/ml) Kit components for 1 box/50 prep Component Column/collection tube Swab/Saliva DNA Solution I Swab/Saliva DNA Solution II DNA Washing Solution I DNA Washing Solution II DNA Elution buffer Product insert Products 50 ea. 6 ml 20 ml 12 ml 12 ml 7.5 ml 1 ea. 5 min Swab/Saliva DNA Extraction Kit
3 5 min Swab/Saliva DNA Extraction Kit Before Starting Add 28 ml of % ethanol to DNA Washing Solution I and II. Mix well. Mark on the labels that ethanol is added. Store both wash buffers with ethanol at room temperature. Procedures for 5 min buccal swab/saliva DNA extraction Important notice; A. All centrifugation in 13,500 rpm. B. Set a Heatblock at 65 o C C. Universal method for DNA extraction of buccal swab, saliva, cervical swab, milk, and semen 1. Add less than 1.0 ml of samples to 1.5 ml of microtube. 2. Microcentrifuge for 30 sec. 3. Remove all supernatant with pipetting or a gentle vacuum. 4. Closed the cap and loosened the pellet by scraping the tube for 5-6 times over an uneven surface such as a microcentrifuge tube rack. 5. Add 100 ul of Swab/Saliva DNA Solution I and vortex for 10 sec. 6. Add 10 ul of Proteinase K (20 mg/ml, not included) and mix well by tapping. 7. Incubate for 15 min at 65 o C. 8. Add 300 ul of Swab/Saliva DNA Solution II and vortex mix for 30 sec. 9. Spin for 5 sec and transfer all lysate to column. 10. Add 350 ul of % isopropanol to column. Close the cap and vortex for 10 sec. 11. Centrifuge for 15 sec. 5 min Swab/Saliva DNA Extraction Kit 3
4 12. Discard the flowthrough. Reassemble the spin column with its collection tube. 13. Apply 700 ul of DNA Washing Solution I and centrifuge for 15 sec. Discard the flowthrough. 14. Apply 400 ul of DNA Washing Solution II and centrifuge for 30 sec. Discard the flowthrough. 15. Replace the collection tube with a clean microcentrifuge tube (not supplied). 16. Add 100 ul of DNA Elution Buffer to column. 17. Close the cap and vortex for 15 sec. 18. Centrifuge for 1 min. 5 min Swab/Saliva DNA Extraction Kit 4
5 Example of test Total DNA extraction from saliva using 5 min Swab/Saliva DNA Extraction Kit. DNA was extracted from 500 ul of 4 different saliva samples and eluted to final 100 ul of elution buffer. Ten ul each DNA was analyzed on 1 % agarose gel. Total DNA extraction from buccal swab using 5 min Swab/Saliva DNA Extraction Kit. DNA was extracted from 500 ul of 6 different buccal swab samples and dissolved to final 100 ul of elution buffer. Ten ul each DNA was analyzed on 1 % agarose gel. 5 min Swab/Saliva DNA Extraction Kit 5
6 Reproducibility of 5 min Sab/ Saliva DNA Kit. Four hundred ul of pooled buccal swab was extracted by the kit and dissolved to final 100 ul of elution buffer. Final 5ul of each sample was analyzed by 1 % agarose gel. Technical Support Biofactories Technical Service Department is staffed by experienced scientists with extensive practical and theoretical expertise in sample and assay technologies and the use of our products. If you have any questions or experience any difficulties regarding products, please do not hesitate to contact us. For technical assistance and more information, please contact our Technical Support Team through at techsupport@5mindna.com. 5 min Swab/Saliva DNA Extraction Kit 6
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Solution WN Wash Solution BE Elution Buffer B Mini Filter Spin Columns 25 Collection Tubes 25 Elution tubes (1.7 ml) 25
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