コントロールセットRNA／DNA
Contents
1.2. 2 DNase RNase 3 RLY CIES DS RO MI K
3. Bim Primer mix H1 NC P PM H1P Positive control H1P PC H1P Negative control
4. 1 Positive control H1P PC H1P A N00 Loopamp RNA DNA BETOR RDO
5. 2 5 8 lt L AMP fait 62 5 C 35 80C 5 95 2 LAMP Positive control H1P PC H1P
6. Negative control NC 1 1 5 0 7 06 0 5 0 4 03 0 2 0 1 0 1 10 20 30 1 JYP gt th 6 CRR VE 4 LAMP
7. ISIEN D F J aR J8 PP oo Loopamp RNA DNA 2 8 C 1 12 LMP248 oe D 3 4 5 Nagamine K et al Molecular and Notomi Nagamine K et al Mori Y
8. 380234 A S to 2015 4 1 opamp LAMP Loop mediated Isothermal Amplification RNA DNA LAMP Loop mediated Isothermal Amplification 1 2 Ds 6 ee ics 4 kN p RNADNA D Loopam JLRS AR 1 Primer mix H1P PM H1P 2 Positive control H1P PC H1P 3 Negative control NC emna ARBER BM gt AS 2 4 A v Loopamp RNA DNA D EH By
9. Essential apparatus equipment and reagents etc Refer to the instruction manual of Loopamp RNA DNA Amplification Reagent D 2 Reagent preparation method follow the procedure of Loopamp RNA DNA Amplification Reagent D for details 1 Take a necessary number total number of samples and controls of the Dried RNA DNA Amplification Reagent Immediately return the remaining reagent to the original aluminum pack and seal it For control reaction this product lt Reagent gt lt Dose gt Primer mix H1P PM H1P 15 0 u L test 2 Mix the reagent PM H1P by inverting tapping or using a vortex mixer for 1 second 3 times and spin down before using as the primer mix Operation on ice 3 Operating procedure Operation on ice Dispense 15 0 u L of PM H1P to each reaction tube Add 10 0 u L of the control to each tube 25 0 uL in total as the LAMP reaction solution Use negative control NC for the negative control and positive control H1P PC H1P for positive control After closing the cap invert the reaction tube to transfer the solution onto the cap and allow it to stand on ice for 2 minutes Repeat the inversion 5 times and spin down the reaction mixture using an 8 microtube simple centrifuge lt LAMP reaction Set it in the real time turbidimeter or incubator reaction block to start reaction The control reaction of this product occurs at 62 5 C for 35 minutes Enzyme d
10. RTS 12 0 72 mL X 1 0 16 mL X 1 0 16 mL X 1 Wh o JU WH gt Primer mix H1P PM H1P 1 X3 RHUL A Loopamp RNADNA D Dried RNA DNA Amplification Reagent gt 15 0uL FAR 1 PM H1P 15 0L 10 0 L LAMP 25 0 uL Negative control NC Positive control H1P PC H1P
11. T et al Nucleic Acids et al Nat Protoc 3 Clin Chem 47 9 Biochem Biophys Res Commun 289 1 Tomita N et al Research 28 12 e63 2000 1742 1743 2001 150 154 2001 5 877 882 2008 Cellular Probes 16 3 223 229 2002 js0120 308 421 A ev 143 For research use only A Loopamp LAMP Loop mediated Isothermal Amplification method Control Set RNA DNA This product is a reagent for research purpose Do not use this product for making or supporting a diagnosis Read this explanatory leaflet carefully before use Introduction The LAMP Loop mediated Isothermal Amplification method is a gene amplification technique characterized by 1 isothermal gene amplification reaction 2 high specificity due to the use of 4 primers recognizing 6 regions 3 high amplification efficiency resulting in amplification in a short time 4 a large amount of amplification product facilitating simple detection This product is a control reaction kit to be combined with Loopamp RNA DNA Amplification Reagent D separately provided by Eiken Chemical Contents For 12 tests 1 Primer mix H1P PM H1P 0 72 mL X 1 2 Positive control H1P PC H1P 0 16 mL X 1 3 Negative control NC 0 16 mL X 1 Instructions for use 1
12. ake care not to directly gaze at the ultraviolet ray sterilizing ray from the lamp of the ultraviolet irradiation device for fluorescent visual evaluation because it is dangerous When it is necessary to gaze at the lamp that is on make sure to do that through a glass plate or using wide eyeglasses or shield 3 A minute amount of sodium azide is contained as a preservative in the primer mix H1P PM H1P positive control H1P PC H1P and negative control NC Take care to prevent sodium azide from entering the eyes or mouth or attaching to skin because it is toxic 4 Ifthe reagent accidentally enters the eyes or mouth or attaches to skin immediately rinse it off with a large amount of water and seek medical treatment if needed Precautions 1 This product should be stored as specified while avoiding freezing or sudden change in temperature 2 Keep the positive control H1P PC H1P away from other reagents 3 Perform gene test with this product only under the supervision of experts with the knowledge and experience of gene test because the lack of knowledge or experience may result in incorrect judgment of the test result 4 Eiken Chemical Co Ltd does not bear any responsibility for false judgment or any consequential damage derived from the false judgment caused by non capability problems such as operation error Use this product within the expiration date Do not recycle the containers or accessories of this product or use t
13. eactivation 80 C for 5 minutes or 95 C for 2 minutes J will be automatically processed in the real time turbidimeter Turbidity measurement evaluation 4 Detection A Real time turbidity detection The target gene can be detected in real time using the real time turbidimeter designed for the LAMP method Refer to the package insert or operation manual etc for the detailed operating procedure B Fluorescent visual detection Judgment is made by emitting ultraviolet ray to the reaction tube from the bottom using the ultraviolet irradiation device and observing the reaction tube from the side with the eyes protected by eyeglasses etc Evaluate the reaction tube according to the following criteria after confirming that the positive control PC H1P produces green fluorescence and the negative control NC produces no fluorescence Take the picture of the reaction tube using a digital camera etc for documentation purposes if required Positive Produces green fluorescence Negative Produces no fluorescence 1 1 5 Amplification curve pattern Positive control Negative control 07 p ja ra 3 0 1 10 20 30 Turbidity Time min Figure 1 Control amplification curve pattern lt Precautions for measurement gt 1 Because the LAMP reaction is very sensitive any contamination with extremely minimum of the target gene or amplification p
14. hem for other purposes 7 Do not use it in combination other regents with the Loopamp RNA DNA Amplification Reagent D Precautions for disposal 1 Appropriately dispose of tubes after reaction with the cap closed by putting them in double plastic bags that can be incinerated or sealed To prevent dispersion of amplification products do not autoclave tubes before disposal 2 The constituent reagent tube is mainly made of polypropylene PP The kit case is mainly made of paper 3 Dispose of this product containers and materials before or after use on the responsibility of the laboratories in compliance with applicable laws on waste disposal and cleaning and water pollution prevention law Storage method shelf life packaging unit and product code ao Product name Storage method Shelf life Package unit Product code Loopamp Control Set 2 8C 1 year For 12 tests LMP248 RNA DNA References 1 Notomi T et al Nucleic Acids Research 28 12 e63 2000 2 Nagamine K et al Clin Chem 47 9 1742 1743 2001 3 Mori Y et al Biochem Biophys Res Commun 289 1 150 154 2001 4 Tomita N et al Nat Protoc 3 5 877 882 2008 5 Nagamine K et al Molecular and Cellular Probes 16 3 223 229 2002 EIKEN CHEMICAL CO LTD Eiken 143 Nogi Nogi machi Shimotsuga gun Tochigi 329 0114 Japan Manufacturer 380234 A Date of Revision April 2015 Ver 1
15. roduct may result in an incorrect result To avoid such contamination the use of this product and specimen collection nucleic acid extraction procedure should be performed in separate rooms or in different areas by partitioning the laboratory area Take appropriate measures to prevent contamination including the use of clean benches gloves and isolation gowns as required 2 Avoid the contamination by microorganisms or nucleic acid degrading enzymes such as DNase and RNase in handling this product 3 Completely dissolve the dry reagent Incomplete dissolution may result in poor performance including low sensitivity Do not leave more than 2 minutes in state of inverting reaction tubes 4 Air bubbles may appear on the liquid level of the reaction solution after mixing the sample solution Remove them to prevent measurement errors by spinning down the reaction mixture 5 Never open the tube cap after reaction Particularly carefully remove the tube from the equipment so as not to open the cap after reaction The contamination with amplification products not only results in an erroneous decision but also causes the contamination of the measurement environment Such contamination may persistently inhibit correct measurement unless it is completely eliminated 6 Avoid handling amplification products using electrophoresis Precautions for handling hazard prevention 1 This product is not designed as an in vitro diagnostic IVD 2 T
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