MagListo 5M Cell Total RNA Extraction Kit

Transcription

1 User s Guide MagListo 5M Cell Total RNA Extraction Kit K-3610 K-3611.

2 MagListo 5M Cell Total RNA Extraction Kit Kit for the extraction of total RNA from wide range of cell types using MagListo User s Guide K-3610 K-3611 Version No.: 3.0 ( ) Please read all the information in booklet before using the unit Bioneer Corporation 8-11, Munpyeongseo-ro, Daedeok-gu, Daejeon 34302, Republic of Korea Tel: Fax: order@bioneer.co.kr MagListo is a trademark of Bioneer Corporation. Copyright Bioneer Corporation. All Rights Reserved.

3 Contents I. Overview 1 II. Kit Components 1 III. Storage 2 IV. Intended Use 2 V. Safety Warnings and Precautions 2 VI. Warranty and Liability 2 VII. Technical Assistance 3 VIII. Quality Management 3 IX. Kit Specifications 4 Extraction of total RNA from small amount of sample 4 X. Sample preparation 5 XI. Principle 5 XII. Magnetic NanoBead Information 6 XIII. Guidelines for MagListo Magnetic Separation Rack 7 XIV. Materials and Equipment Needed But Not Provided 7 Types of the Magnetic Separation Rack 8 XV. Procedure 9 XVI. Protocols 10 Before you begin 10 A. RNA Extraction from Cultured Cells 10 B. RNA Clean up (RNA Purification) 15 C. ONE Step RNA Cleanup (DNase Treatment) 16 XVII. Appendix 18

4 Troubleshooting guide 18 Experimental data 20 XVIII. Ordering Information 22 XIX. Explanation of Symbols 23

5 I. Overview Description MagListo 5M Cell Total RNA Extraction Kit utilizes Magnetic Nano Beads to extract total RNA from cultured cells, with the aid of the MagListo Magnetic Separation Rack. The use of MagListo Magnetic Separation Rack along with this kit greatly increases user convenience by reducing the total RNA extraction time without centrifugation. Features and Benefits - Magnetic Nano Beads enable the rapid nucleic acid extraction - No requirement of expensive instruments - A single kit serves mini or midi scale experiment Applications Applicable to assays requiring RNA, including, but not limited, RT-PCR, cdna synthesis, Northern, dot, and slot blot analyses, Primer extension, Poly A + RNA selection, Microarrays II. Kit Components MagListo 5M Cell Total RNA Extraction Kit * K-3611 Buffer 1 (Binding) 25 ml x 2 ea Buffer 2 (1 st Washing) 80 ml x 1 ea Buffer 3 (2 nd Washing) 80 ml x 1 ea Buffer 4 (3 rd Washing) 120 ml x 1 ea Buffer 5 (Elution) 25 ml x 1 ea Magnetic Nano Bead - RNA 1.8 ml x 6 ea *mini 100 rxn, midi 20 rxn 1

6 III. Storage MagListo 5M Cell Total RNA Extraction Kit should be stored dry at room temperature. It can be stored for up to 2 years if it remains sealed. IV. Intended Use MagListo 5M Cell Total RNA Extraction Kit is intended for research use only. This kit is not intended for human or veterinary diagnostics. V. Safety Warnings and Precautions Please inquire BIONEER s Customer Service Center to obtain a copy of the Material Safety Data Sheet (MSDS) for this product. Before, during and after using this kit, all potentially hazardous materials (i.e. materials that may have come in contacted with genetically recombinant samples) including tubes, tips and other kit contents should be processed and discarded in accordance with applicable and appropriate regulations of the municipality/government in which this product is being used. A user must also be equipped with basic experimental techniques required for correct execution of the extraction experiments described in this User s Guide. Some inventions applied in this kit may infringe existing patents in certain countries. The purchase of this kit does not include or provide a license to perform such patented inventions. Users may be required to obtain a license depending on the patent law of the country where this product is being used. We do not condone nor recommend the unlicensed use of patented inventions. VI. Warranty and Liability All BIONEER products are manufactured and tested under strict quality control protocols. BIONEER guarantees the quality of all directly manufactured products during the warranty period of one (1) year from the date of purchase. If you find any issues regarding the product quality, please immediately contact BIONEER s Customer Service Center (sales@bioneer.com). BIONEER does not assume any liability for the misuse of the product, i.e. using the product for any purposes other than its intended purpose as described in the User s Guide. BIONEER will only assume 2

7 liability under the condition when the users disclose all related information regarding the issue to BIONEER in written form within 30 days after occurrence of the issue. VII. Technical Assistance At Bioneer, we pride ourselves on being responsive to your needs. Our Technical Service Departments are staffed by experienced scientists with extensive practical and theoretical expertise in Molecular Biology and the use of Bioneer products. If you have any questions or would like to find out more information about MagListo products, please contact us. We look forward to hearing from you! Technical Support For all technical questions and troubleshooting on Bioneer products and applications Tel: In North America Tel: VIII. Quality Management Every aspect of our quality management system from product development to supplier qualification ensures that our products meet the world-class standards. Each lot of MagListo 5M Cell Total RNA Extraction Kit is carefully tested by the quality control team. 3

8 IX. Kit Specifications Micro scale Mini scale Midi scale Starting Cell Number x x x 10 7 Extraction time < 8 min < 8 min < 13 min Elution volume µl µl µl Expected RNA yield Up to 1 µg Up to 100 µg Up to 400 µg Expected purity A 260/ 280 > 1.9 *RNA content can vary greatly among cell types. Typical RNA yield from cultured cell purified with MagListo 5M Cell Total RNA Extraction Kit. Cell type (1x 10 6 ) Yield Hela µg 293T µg Balb/c 3T µg Huh µg Extraction of total RNA from small amount of sample MagListo 5M Cell Total RNA Extraction Kit is also able to extract total RNA from a small quantity of sample. Refer to Cell Total RNA Extraction from Cultured Cell for Micro in page 10 for more details about RNA extraction from samples with a low number of cells (< 1 x 10 4 ). 4

9 Table1. Growth area and Average cell yield in various culture dishes. Cell culture dishes Growth area (cm 2 ) Average cell yield Multi well plate 6 well x well 4 4 x well 2 2 x well 1 1 x well x 10 4 Dishes 35 mm x mm 21 3 x mm 55 8 x mm x 10 7 Flasks 50 ml x ml 75 1 x 10 7 X. Sample Preparation Cultured cells can easily be harvested using a centrifuge. However, it might be difficult to extract total RNA if cultured cells are too clustered. In this case, trypsin can be used to detach clustered cells. For extraction, the number of cells should be less than 1 x 10 7, which is calculated with a cell counter. It is recommended to keep samples on ice before use. Xl. Principle The MagListo 5M Cell Total RNA Extraction Kit is designed for the extraction of high purified total RNA from cultured cells. The overall principle is based on adsorption of RNA onto the Magnetic Nano Bead by chaotropic salt. For example, chaotropic agents in Buffer 1 (Binding) contains guanidine hydrochloride and guanidine thiocyanate, as which remove water molecules around RNA and silica coated magnetic beads surface resulting in RNA then being captured by magnetic beads. The Magnetic Nano Beads and RNA complexes are pulled and fixed on the tube wall using a magnetic force, followed by washing with ethanol to remove debris and excessive salts. Finally, the captured RNAs are then eluted by Buffer 5 (Elution), an aqueous solution with optimal ph. 5

10 Lysis Binding Washing Elution XII. Magnetic Nano Bead Information Description Magnetic Nano Beads have been developed to overcome shortcomings of existing resin and to automate purification process. The principle of extraction using the Magnetic Nano Beads is that coated functional group on the surface of the Magnetic Nano Beads bind with DNA and the Magnetic Nano Beads are then isolated using external magnetic field. Features Fast binding guarantees higher throughput automation Large Surface Area enables more sensitive assay Globular structure increases specificity by decreasing non-specific binding Specification Matrix Silica-coated Fe 3O 4 AccuNanoBead Silica Magnetic Nano Beads Average size Ligand 400nm -OH Working Temp Storage Store at room temperature upon receipt 6

11 XIII. Guidelines for MagListo Magnetic Separation Rack Description MagListo Magnetic Separation Rack is designed for a fast, easy separation of the Magnetic Nano Beads. These racks of different sizes allow users to choose the product according to their needs. The following are recommended when handling the MagListo Magnetic Separation Rack The product is made of acryl and plastic. Be careful not to drop the product as the dropping may break the product. When moving the product, take extra care not to drop the product as it may cause injury. If the product is broken, do not discard it with bare hands as the sharp edges may cause injury. When an extracted or purified nucleic acid is spilled on the product, immediately rinse it with running water and clean it with 70% ethanol. Acetone, Toluene, or other organic solvent may damage the acrylic and plastic part of the product, which may lead to malfunction of the product. Rinse the product immediately when spillage of any above mentioned solvents occurs as the expected DNA yield may not be obtained if the product is damaged. Make sure that a corrosive liquid does not spill on the magnet plate part of the product. If spillage occurs, immediately rinse it off with running water as it may corrode the magnet during storage and may degrade its performance. XIV. Materials and Equipment Needed But Not Provided 1. Table-top microcentrifuge, 16,000 x g (>13,000 rpm) (mini scale) 2. Centrifuge with rotor capable of 3,000 x g (midi) ml or 2 ml tube (micro / mini scale) ml tube (midi scale) 5. Vortex mixer 6. Absolute ethanol 7. Thermal block 8. MagListo Magnetic Separation Rack 7

12 Types of the Magnetic Separation Rack Tube MagListo Magnetic Separation Rack Cat.No 1 ml tube with 8-cap strip MagListo -8Ch Magnetic Separation Rack TM ml or 2 ml microcentrifuge tube MagListo -2 Magnetic Separation Rack TM ml tube MagListo -15 Magnetic Separation Rack TM-1020 (Note) Please refer to the ordering information in this User s Guide for more information regarding catalog number of racks designed for specific size of tubes. 8

13 XV. Procedure-Cell Total RNA Extraction 9

14 XVI. Protocols Before you begin 1. Buffer 1 (Binding) and Buffer 2 (1 st Washing) contains chaotropic salt. You should take the appropriate laboratory safety precautions and wear gloves and lab goggle when handling. 2. The relative centrifugal force (RCF) is calculated in g as follows: RCF = 1.12 x r x (rpm/1,000) 2 Where r is the radius of a rotor in cm, and rpm is the speed of the rotor in revolutions per minute. 3. To inhibit RNase activity, we recommend adding β-mercaptoethanol to Buffer 1 (Binding) before use. Add 10 µl β-mercaptoehanol(>99%) per 1 ml Buffer 1 (Binding). A. RNA Extraction from cultured cell for Micro/Mini/Midi scale 1. (Harvest cell) Cells grown in suspension : Count the cell number and centrifuge given number of cells (~1x10 4 (micro) / ~5x10 6 (mini) / ~2x10 7 (midi)) at 300 x g for 5 min. Discard supernatant carefully and go to lysis & homogenization (go to step 3). 2. Cells grown in a monolayer: There are 2 different ways to collect cells grown in a monolayer. a. Direct cell lysis on the culture dish: Completely remove Cell Culture Medium and go to lysis & homogenization (go to step 3). (Remaining medium may inhibit the RNA extraction) b. Harvesting cells with trypsin: Remove Cell Culture Medium and wash the monolayer with DPBS. Add 0.1%-0.25% typsin to the washed cell monolayer. When the cells are detached, add Cell Culture Medium to inactivate the typsin. Transfer the cells into a RNase-free tube and centrifuge at 300 xg for 5 min. Discard supernatant carefully and go to lysis & homogenization (go to step 3). 3. (Lysis & homogenization) Add 50 µl (micro) / 400 µl (mini) / 2 ml (midi) of Buffer 1 (Binding) to each sample and mix thoroughly using a vortex mixer. Make sure that you must completely resuspend the sample to achieve maximum lysis efficiency. (Note) Insufficient homogenization can decrease the RNA purification yield, and also cause 10

15 clogging of Magnetic Nano Beads in the following steps. For the sufficient homogenization of the lysate, make the lysate passed through blunt 20-gauge needle (0.9 mm diameter) 5 to 10 times. A. (Micro / Mini) please transfer the lysate to a 1.5 ml or 2 ml tube. B. (Midi) please transfer the lysate to a 15 ml tube. 4. (RNA precipitation) Add 50 µl (micro) / 400 µl (mini) / 2 ml (midi) of absolute ethanol to the each tube and mix well using a vortex mixer or by pipetting. 5. (RNA binding with Magnetic Nano Bead: 5-7) Add 50 µl (micro) / 100 µl (mini) / 400 µl (midi) of Magnetic Nano Bead solution to the tube and mix thoroughly using a vortex mixer until the beads are fully resuspended. (Note) Magnetic Nano Bead solution contains Magnetic Nano Beads. Please shake well or mix with a vortex mixer before use. 6. Place the tubes on the MagListo -2 (micro, mini) / MagListo -15 (midi) Magnetic Separation Rack with the magnet plate attached and invert the rack gently 3 to 4 times until the beads bind tightly to the magnet. - Attachment of the magnet plate Combine the magnet plate to the stand. 7. Without removing the tube from MagListo Magnetic Separation Rack, discard the supernatant carefully and completely remove the remaining supernatant on a paper towel action. - How to discard the supernatant 11

16 - Discard the supernatant by inverting the MagListo Magnetic Separation Rack. The silicone immobilizer inside the stand prevents the tubes from falling. When discarding the supernatant, invert the rack completely so that the solution does not to spill on the rack. 8. (1 st washing: 8-10) Detach the magnet plate from MagListo Magnetic Separation Rack. Add 350 µl (micro) / 700 µl (mini) / 3.5 ml (midi) of Buffer 2 (1 st Washing) to the each tube and close the cap. Mix by vortexing or shaking until the beads are fully resuspended. -Detachment of the magnet plate Detach the magnet plate gently by pulling it upwards. 9. Attach the magnet plate and invert the rack gently 3 to 4 times until the beads bind tightly to the magnet. 10. Without removing the tubes from the MagListo Magnetic Separation Rack, discard the supernatant and remove the remaining supernatant on a paper towel by blotting action. 11. (2 nd washing) Repeat the steps 8-10 by adding 350 µl (micro) / 700 µl (mini) / 3.5 ml (midi) of Buffer 3 (2 nd Washing) instead Buffer 2 for additional washing. 12. (3 rd washing) Without removing the tubes from the MagListo Magnetic Separation Rack, add 700 μl (micro, mini) / 6 ml (midi) of Buffer 4 (3 rd Washing) to the opposite side of bead pellet. Close the cap and gently invert the rack twice in order to remove ethanol from the sample. (Note) Direct pipetting of Buffer 4 onto the bead pellet, vortexing and/or vigorous shaking of the tubes may release nucleic acid from the beads, which may result in lower RNA yield than expected. 12

17 13. Discard the supernatant and completely remove the remaining supernatant by blotting action. - Add Buffer 4 and discard the supernatant 14. (Elution: 14-18) Detach the magnet plate from the MagListo Magnetic Separation Rack. Add µl (micro, mini) / µl (midi) of Buffer 5 (Elution) to the tube with the magnet plate detached and resuspend completely by pipetting or vortex mixer for 15 sec. 15. Incubate the tube at for 1 min. 16. Place the tubes on the MagListo Magnetic Separation Rack. Attach the magnet plate and invert the rack gently 3 to 4 times until the beads bind tightly to the magnet. 17. Without removing the tubes from the MagListo Magnetic Separation Rack, transfer the supernatant containing RNA to a new sterile microcentrifuge tube. 18. Discard the tubes with remaining Magnetic Nano Bead pellet. Do not reuse the beads. 13

18 Summary of reagent volumes required in each step of Cell Total RNA Extraction Step Buffer Micro Mini Midi Page Lysis Buffer 1 (Binding) 50 μl 400 μl 2 ml P. 10 RNA Precipitation Absolute Ethanol 50 μl 400 μl 2 ml P. 11 Bead Binding Magnetic Nano Bead 50 μl 100 μl 400 μl P st Washing Buffer 2 (1 st Washing) 350 μl 700 μl 3.5 ml P nd Washing Buffer 3 (2 nd Washing) 350 μl 700 μl 3.5 ml P rd Washing Buffer 4 (3 rd Washing) 350 μl 700 μl 6 ml P. 12 Elution Buffer 5 (Elution) μl μl ul P

19 B. RNA Clean up (RNA Purification) 1. Transfer the eluted RNA or enzyme reaction product to a new sterile tube. A. (Micro / Mini) Transfer the eluate to a 1.5 ml or 2 ml tube B. (Midi) Transfer the eluate to a 15 ml tube 2. (Binding) Add 1 volume of Buffer 1 (Binding) to 1 volume of the eluted RNA and mix completely using a vortex mixer. 3. (RNA precipitation) Add 2 volumes of absolute ethanol to 1 volume of the eluted RNA and mix completely using a vortex mixer. 4. (RNA binding with Magnetic Nano Bead: 5-7) Add 50 µl (micro) / 100 µl (mini) / 400 µl (midi) of Magnetic Nano Bead solution to each tube and mix thoroughly using a vortex mixer until the beads are fully resuspended. (Note) Magnetic Nano Bead solution contains Magnetic Nano Beads. Please shake well or mix with a vortex mixer before use. 5. Place the tubes on the MagListo -2 (micro, mini) / MagListo -15 (midi) Magnetic Separation Rack with the magnet plate attached and invert the rack gently 3 to 4 times until the beads bind tightly to magnet. 6. Without removing the tube from MagListo rack, carefully pour the supernatant out and completely remove the remaining supernatant using paper towel by blotting action. 7. Go to step 8 of A. RNA Extraction from Cultured Cell in page 12 and follow the instructions accordingly. 15

20 C. One Step RNA Clean Up (DNase Treatment) 1. (RNA precipitation) Detach the magnet plate from the MagListo Magnetic Separation Rack. Add 400 µl (micro, mini) / 3.5 ml (midi) of absolute ethanol to the each tube and close the cap. Mix by vortexing or shaking until the bead are fully resuspended 2. Place the tubes on MagListo -2 (mirco, mini) / MagListo -15 (midi) Magnetic Separation Rack with the magnet plate and invert the rack gently 3 to 4 times until the beads bind tightly to the magnet. 3. Without removing the tubes from the MagListo TM Magnetic Separation Rack, discard the supernatant carefully and completely remove the remaining supernatant using a paper towel by blotting action. 4. The beads can be dried with a dry oven at 65 for following times. (micro/mini : >5 min, midi : >25 min) Please use a clean bench during the drying procedure to prevent RNase or other aerosol contamination. 5. Add 50 μl of DNase Reaction Buffer and 10 μl of RNase-Free DNaseⅠ to the each tube. 6. Detach the magnet plate from the MagListo Magnetic Separation Rack and close the cap. Mix using a vortex mixer until the beads are fully resuspended. 7. Place on the benchtop (20-30 C) for 20 min. 8. Detach the magnet plate from MagListo Magnetic Separation Rack. Add 350 µl (micro) / 700 µl (mini) / 3.5 ml (midi) of Buffer 2 (1 st Washing) to the each tube and close the cap. Mix using a vortex mixer until the beads are fully resuspended. 9. Place the tubes on the MagListo Magnetic Separation Rack with the magnet plate and invert the rack gently 3 to 4 times until the beads bind tightly to the magnet. 16

21 10. Without removing the tubes from the MagListo Magnetic Separation Rack, discard the supernatant carefully and completely remove the remaining supernatant using a paper towel by blotting action. 11. Go to step 11 of A. RNA Extraction from Cultured Cell in page 12 and follow the instructions accordingly. 17

22 XV. Appendix Troubleshooting guide This troubleshooting guide will help you to solve problem that may arise during RNA extraction. For other technical assistance or more information, please contact our technical assistance team. Comments and suggestions Buffers or other reagents may have been exposed to external factors that may have reduced its quality. Please make sure that reagents are stored at room temperature at all times upon arrival and that all reagent bottles are closed tightly, in order to preserve ph and stability, and to avoid contamination. Too much amount of starting sample was used to extract RNA. Appropriate amount of starting sample (see Kit Specification in page 4) should be used. Elution may have been incomplete. Please extend incubation time up to 3 minutes at elution step to improve the yield. In addition, make sure that Low yield of RNA Magnetic Nano Beads are resuspended completely in the eluting solution during incubation. Some of Magnetic Nano Bead pellet may have been lost while discarding solution. Check that all of the Nano Beads have bound tightly to the magnet when you discard supernatant. Insufficient shaking or vortexing during lysis step may lead to low RNA yield than expected. Shake or mix the cell lysate with a vortex mixer sufficiently during incubation step. Cell culture medium may have been removed incomplete. Try to remove the medium as much as possible. Any leftover in the medium can lead to the inhibition of RNA extraction. 18

23 Beads may have been washed insufficiently. You must properly wash the beads in the 3 rd washing step. Remaining ethanol can decrease the purity of Low A 260/280 ratio RNA. Take enough time to wash the beads properly. Incomplete suspension of beads during the washing step causes salts to remain in the purified RNA. Make sure that the beads are suspended thoroughly during the washing process. Excessively clustered Magnetic Nano Bead Excess amount of starting sample is used to extract RNA. Appropriate amount of starting material (see Kit Specification in page 4) should be used for efficient extraction of RNA. Presence of a white precipitate in buffers A white precipitate may form in Buffer l (Binding) due to prolonged storage at low temperatures. Incubate Buffer l (Binding) at 60 until the precipitate to be dissolved. RNase contamination can be degraded RNA. Use a heat gun or a blow dryer in a clean bench to prevent the contamination of RNase in the air. Use Degraded RNA RNase-free pipette tips and change the gloves frequently. Frequent freezing and thawing may result in lower RNA yield than expected. Avoid repeated freezing and thawing. Flotation of extracted RNA floats when loaded on an agarose gel Floating of RNA on an agarose gel is caused by the remaining ethanol in the eluted RNA. Ensure that the 3 rd Washing (ethanol removing) step in the protocol is properly performed. Remaining ethanol may also interrupt the enzymatic reaction. 19

24 Experimental Data Figure 1: Comparison of total RNA purified with MagListo 5M Cell Total RNA Extraction Kit and competitor 1x10 6 and 5x10 6 Hela cells were used for the comparison. Total RNA purification was performed with the MagListo 5M Cell Total RNA Extraction Kit and the competitor s product respectively. The bands of purified total RNA were illustrated by 1% denaturing agarose gel electrophoresis. The equivalent level of purification yield of the MagListo 5M Cell Total RNA Extraction Kit and the competitor s product was confirmed through the bands. Also the distinctive 28S/18S rrna band patterns represent the superb quality of RNA purities having no RNA degradations. Figure 2: Extraction of RNA from various cell lines 1X10 6 of Huh7, Hela, 293T, and Balb/c 3T3 cells were used for the RNA purification performed with the MagListo 5M Cell Total RNA Extraction Kit. The figure shows the bands of purified RNA made by using 1% denaturing agarose gel electrophoresis. The effective RNA purifications from various cell lines could be confirmed through the bands. 20

25 Ct value MagListo 5M Cell Total RNA Extraction Kit Figure 3: RT-qPCR comparison of RNA isolated from cells Hela Cell-GAPDH MagListo Competitor R² = R² = E+06 1.E+05 1.E+04 1.E+03 1.E+02 1.E+01 Cell Number (Left) Human GAPDH extracted from 10, 10 2, 10 3, 10 4,10 5, 10 6 Hela cells with the MagListo 5M Cell Total RNA Extraction Kit were amplified using the AccuPower RoketScript RT-qPCR Premix (K-6700) kit. The figure represents fluorescent signals of amplified Human GAPDH. (Right) The graph shows the comparison results of amplified Hela cell-gapdh extracted with the MagListo 5M Cell Total RNA Extraction Kit and the competitor s product. According to the results, RNAs were successfully purified from 10~10 6 Hela cells. Also the equivalent level of purification yield of the MagListo kit and the competitor s product was confirmed through the GAPDH Ct value of each cell number. 21

26 XIV. Ordering Information Cat no. Product Description Size K-3601 MagListo TM 5M Plamid Extraction Kit, 100 reactions (mini) 1 kit K-3600 MagListo TM 5M Plamid Extraction Kit, 500 reactions (mini) 1 kit K-3603 MagListo TM 5M Genomic DNA Extraction Kit, 100 reactions (mini) 1 kit K-3605 MagListo TM 5M Plant Genomic DNA Extraction Kit, 100 reactions (mini) 1 kit K-3607 MagListo TM 5M Gel Extraction Kit, 100 reactions (mini) 1 kit K-3609 MagListo TM 5M PCR Purification Kit, 100 reactions (mini) 1 kit K-3611 MagListo TM 5M Cell Total RNA Extraction Kit, 100 reactions (mini) 1 kit K-3613 MagListo TM 5M Tissue Total RNA Extraction Kit, 100 reactions (mini) 1 kit K-3615 MagListo TM 5M Forensic Sample DNA Extraction Kit, 100 reactions (mini) 1 kit K-3617 MagListo TM 5M Viral DNA / RNA Extraction Kit, 100 reactions (mini) 1 kit K-3619 MagListo TM 5M Circulating Cell Free DNA Extraction Kit, 50 reactions (mini) 1 kit TM-1000 MagListo TM -8Ch Magnetic Separation Rack 1 ml tube x 8 holes TM-1010 MagListo TM -2 Magnetic Separation Rack 2 ml tube x 8 holes TM-1020 MagListo TM -15 Magnetic Separation Rack 15 ml tube x 6 holes TM-1030 MagListo TM -50 Magnetic Separation Rack 50 ml tube x 3 holes HT-15-NG 1.5 ml microcentrifuge tube 500 ea / pk HT-20-NG 2 ml microcentrifuge tube 500 ea / pk 22

27 XIX. Explanation Symbols Catalog Number Batch code Manufacturer Contains sufficient for (n) tests Caution, consult accompanying documents Caution, Potential Biohazard USE BY Temperature Limitation DO NOT REUSE Consult Instruction For Use 23

28 8-11 Munpyeongseo-ro, Daedeok-gu, Daejeon, 34302, Republic of Korea (Korea: ) Marina Village PKWY, Suite 110, Alameda, CA 94501, USA (Toll-free) us.bioneer.com Korea Bio Park BLDG #B-702, 700 Daewangpangyo-ro, Bundang-gu, Seongnam-si Gyeonggi-do, 13488, Republic of Korea