5 min Cell/Virus DNA/RNA Extraction Kit
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1 5 min Cell/Virus DNA/RNA Extraction Kit Catalog #; N-1002 The fastest and the simplest but most reliable DNA/RNA extraction system Co-purification of DNA/RNA from animal cell, bacteria, fungi, yeast, protozoa, and blood. Co-purification of viral DNA/RNA from serum, plasma, CSF, wash, culture media, saliva, buccal swab, nasal swab, and throat swab. Total procedure in 5 minutes An ideal tool for both DNA and RNA extraction from a limited sample No damage to RNA quality during extraction procedure Every procedure at ambient temperature without any cold or freezing step Single column format Final 5-20 ug of total DNA/RNA The kit can be stored at ambient temperature for more than a year. One kit for 50 purification The extracted DNA/RNA can be used in any downstream steps without any inhibitory effect. 5 min Cell/virus DNA/RNA 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 % isopropanol % ethanol Sterile cotton swab and scissors (for nasal and throat swab preparation) Kit components for 1 box/50 prep Component Column/collection tubes Cell/Virus DNA/RNA Solution DNA/RNA Washing Solution Elution Solution Product insert Products 50 ea 30 ml 18 ml 7.5 ml 1 ea 5 min Cell/Virus DNA/RNA Extraction Kit
3 5 min Cell/Virus DNA/RNA Extraction Kit Before Starting Add 42 ml of % ethanol to Cell/Virus DNA/RNA Washing Solution. Mix well. Mark on the labels that ethanol is added. Store it at room temperature. Procedures for lysate preparation for Cell/Virus DNA/RNA extraction Notice; All centrifugation in 10,000 rpm at room temperature. All steps at ambient temperature A. Cultured animal cells General notice for lysate preparation 1. The maximum recommended input of cells is 2.5 x As a reference, confluent 60 mm and 100 mm dish contain 3.2 X10 6 and 8.8 X10 6 cells respectively. 2. The lysate can be stored at -80 o C for later RNA extraction. 3. For DNA/RNA extraction from frozen cell, loosened the pellet by scraping the tube for 5-6 times over an uneven surface such as a microcentrifuge tube rack and add lysis solution directly to frozen sample. Lysate preparation Cell grown in monolayer 1. Aspirate media 2. Add 350 ul of Cell/Virus DNA/RNA Solution and mix with gentle swirling for 30 sec. 3. Transfer all lysate to a microtube and vortex for 15 sec. 4. Add 350 ul of isopropanol and vortex for 10 sec. 5 min Cell/Virus DNA/RNA Extraction Kit 3
4 Cell grown in suspension or detached monolayer cells 1. Transfer cell suspension to a microtube (not provided) and centrifuge for 30 sec. 2. Remove supernatant with pipetting or a gentle vacuum. 3. Repeat steps 1 and 2 as required. 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 350 ul of Cell/Virus DNA/RNA Solution and vortex for 15 sec. 6. Add 350 ul of isopropanol and vortex for 10 sec. B. Bacteria, fungi, yeast, and protozoa General notice for lysate preparation 1. It is recommended that no more than 1.5 ml of saturated bacterial or yeast culture volume be used in this procedure in order to prevent clogging of the column. 2. It is recommended that no more than 100 mg of fungi or protozoa be used for this procedure in order to prevent clogging of the column. Lysate preparation 1. Transfer cell in culture media or PBS to a microtube (not provided) and centrifuge for 30 sec. 2. Remove supernatant with pipetting or a gentle vacuum. 3. 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. 4. Add 350 ul of Cell/Virus DNA/RNA Solution and vortex for 30 sec. 5. Add 350 ul of isopropanol and vortex for 10 sec. C. Nasal or Throat Swabs Lysate preparation 1. Transfer 350 ul of Cell/Virus DNA/RNA Solution to a clean microtube (not provided) 2. Gently brush a sterile, single-use cotton swab (not provided) inside the nose or mouth of the subject. 3. Cut the cotton tip of swab with clean scissors and leave in the microtube. 5 min Cell/Virus DNA/RNA Extraction Kit 4
5 4. Closed the cap and vortex for 30 sec. 5. Add 350 ul of isopropanol and vortex for 10 sec. D. Saliva and Buccal Swabs Lysate preparation 1. Transfer specimen to a microtube (not provided) and centrifuge for 30 sec. 2. Remove supernatant with pipetting or a gentle vacuum. 3. 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. 4. Add 350 ul of Cell/Virus DNA/RNA Solution and vortex for 15 sec. 5. Add 350 ul of isopropanol and vortex for 10 sec. E. Viral RNA from serum, plasma, CSF, wash, and media Lysate preparation for viral RNA 1. Transfer less than 0.3 ml of sample to a microtube 2. Add 0.6 ml of Cell/Virus DNA/RNA Solution and vortex mix for 30 sec. 3. Add 0.6 ml of isopropanol and vortex for 10 sec. 5 min Cell/Virus DNA/RNA Extraction Kit 5
6 Procedures for Cell/Virus DNA/RNA Extraction from the lysate Important notice; A. All centrifugation in 10,000 rpm at room temperature. B. If column is clogged, spin more until a complete flowthrough. A. Universal procedure for all lysate B. All steps at ambient temperature 1. Transfer all lysate to column. Do not centrifuge to collect the sample from the lid. Any centrifuge in this step would severely reduce the amount of purified DNA/RNA. 2. Centrifuge the column for 15 sec. 3. Discard the flowthrough. Reassemble the spin column with its collection tube. 4. Repeat the steps of 1 and 3 if required. 5. Apply 700 ul of DNA/RNA Washing Solution and centrifuge for 15 sec. Discard the flowthrough. 6. Apply 400 ul of DNA/RNA Washing Solution and centrifuge for 30 sec. Discard the flowthrough. 7. Replace the collection tube with a clean microcentrifuge tube (not supplied). 8. Add 100 ul of DNA/RNA Elution Buffer to column. 9. Close the cap and vortex for 15 sec. 10. Centrifuge for 15 sec. 11. Reload the flowthrough to the top of column and centrifuge for 30 sec. 5 min Cell/Virus DNA/RNA Extraction Kit 6
7 Example of test Total DNA/RNA extraction by 5 min Cell/Virus DNA/RNA Extraction Kit. DNA/RNA was extracted from 1 ml of 1 week old Saccharomyces cerevisiae culture followed by the supplied protocol. Total DNA/RNA was eluted to final 100 ul of Elution Buffer and analyzed on 1 % agarose gel by 10 ul each. Total DNA/RNA extraction from cultured cells using 5 min Cell/Virus DNA/RNA Extraction Kit. DNA/RNA was extracted from each of 2 X10 6 of indicated cultured cells followed by the supplied protocol. Total DNA/RNA was eluted to final 100 ul of Elution Buffer and analyzed on 1 % agarose gel by 5 ul each. Lanes 1 to 3; HeLa cell, lanes 4 to 6; HEK 293 cells. Total DNA/RNA extraction from saliva and buccal swab using 5 min Cell/Virus DNA/RNA 5 min Cell/Virus DNA/RNA Extraction Kit 7
8 Extraction Kit. DNA/RNA was extracted from each of 100 mg of indicated tissue followed by the supplied protocol. Total DNA/RNA was eluted to final 100 ul of Elution Buffer and analyzed on 1 % agarose gel by 5 ul each. Lanes 1 to 3; buccal swab, lanes 4 to 6; saliva. Total DNA/RNA extraction from E. coli using 5 min Cell/Virus DNA/RNA Extraction Kit. DNA/RNA was extracted from 1.5 ml of fresh grown saturated culture followed by the supplied protocol. Total DNA/RNA was eluted to final 100 ul of Elution Buffer and analyzed on 1 % agarose gel by 5 ul. Co-purification of DNA and RNA by 5 min Cell/Virus DNA/RNA Kit. Two hundred fifty ul of HBV or HCV positive plasma samples were used for DNA/RNA extraction by 5 min Cell/Virus DNA Extraction Kit (DNA extraction), 5 min Cell/Virus RNA Extraction Kit (RNA extraction), or 5 min Cell/Virus DNA/RNA Extraction Kit (DNA/RNA extraction). Each extracted DNA or RNA was used for qpcr (for HBV) or qrt-pcr (for HCV) by CFX96 (BioRad) in the presence of each 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 Cell/Virus DNA/RNA Extraction Kit 8
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3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Phage DNA Isolation Kit Product # 46800, 46850 Product Insert
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3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Water RNA/DNA Purification Kit Product # 26480 Product Insert
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3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Food DNA Isolation Kit Product # 54500 Product Insert Norgen s
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3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Plant/Fungi Total RNA Purification Kit Product # 25800, 31350,
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3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Norovirus RT-PCR Detection Kit Product # 41400 Product Insert
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3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Plant/Fungi DNA Isolation Kit Product # 26200, 26250 Product Insert
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3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Canine Parvovirus PCR Detection Kit Product # 45000 Product Insert
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3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Size-Select Kit for NGS Library Preparation Product # 53600 Product
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3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com DNA Gel Extraction Kit Product # 13100 Product Insert The Norgen