5 min Stool DNA Extraction Kit

Transcription

1 5 min Stool DNA Extraction Kit Catalog #; D-1003 The fastest and simplest but most reliable Nucleic Acid extraction system Purification of total DNA from host and microorganisms in the stool/feces. Total procedure in 5 minutes without any hidden steps Minimum hands on time Single column format Final 4-15 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 Stool 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 Kit components for 1 box/50 prep Component Column/collection tube Stool DNA Solution I Stool DNA Solution II DNA Washing Solution I DNA Washing Solution II DNA Elution buffer Product insert Products 50 ea. 40 ml 30 ml 12 ml 12 ml 7.5 ml 1 ea. 5 min Stool DNA Extraction Kit

3 5 min Stool 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 DNA extraction from stool/feces Important notice; A. All centrifugation in 13,500 rpm. B. Frozen or fresh stool and feces can be used. 1. Transfer less than 0.3 g of fresh or frozen stool/feces samples to 1.5 ml microtube 2. Break the solid part of stool/faces using a 1 ml pipette tip. 3. Add 700 ul of Stool DNA Solution I and vortex for 30 sec. 5. Centrifuge only for 2-3 seconds. With more centrifugation, the yield for DNA can be decreased in the final step. 6. Transfer 600 ul of solution mixture to 1.5 ml microtube 7. Spin for 30 sec. and remove all liquid by pipetting or gentle vacuum. 8. 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. 9. Add 400 ul of Stool DNA Solution II and vortex for 30 sec. 10. Spin 15 sec. and load all lysate to column. 11. Add 300 ul of isopropanol to the column and vortex mix for 10 sec after closing of the cap. 12. Spin for 15 sec. and discard the flowthrough. 13. Add 700 ul of DNA Washing Solution I and spin for 15 5 min Stool DNA Extraction Kit 3

4 14. Add 400 ul of for DNA Washing Solution II and spin for 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. Example of test Total DNA extraction from six different stool samples using 5 min Stool DNA Extraction Kit. DNA was extracted based on the supplied protocol and eluted to 100 ul of Elution buffer. Ten ul each DNA was analyzed on 0.8 % agarose gel. 5 min Stool DNA Extraction Kit 4

5 There is no PCR inhibitory molecules in stool DNA purified by 5 min Stool DNA Extraction Kit. The stool DNA purified by previous Figure was used for PCR reaction without any dilution (1 X samples) or after 10 fold dilution (1/10 samples) in the presence of hactin specific primers. 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 Stool DNA Extraction Kit 5