Safety information - Eurogentec

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2 Table of contents Intended use 3 Shipping and storage conditions 3 Safety information 3 Background Information 4 Kit contents 6 MiRmaid miRNA precursors qRT-PCR primer set components 7 Additional required material 7 Standard protocol 8 1. Reagent handling 8 2. RNA preparation 8 3. DNase and Reverse Transcription Reactions 9 4. Quantitative qPCR Reaction 10 5. How to set up a good assay 12 Typical results 14 Troubleshooting guide 15 Homology between human pre-miRNA primers and rodent sequences 19 Guidelines for successful RT-qPCR 20 Ordering information 21 Related products 23 Appendix 1: Layout of MiRmaid qPCR primer plate #1 24 Appendix 2: Layout of MiRmaid qPCR primer plate #2 24 Appendix 3: qPCR MasterMixes compatibility with Real-Time thermocyclers 25 Trademarks and disclaimer 26 Manual for MiRmaid miRNA precursor qRT-PCR Primer Set I www.eurogentec.com I info@eurogentec.com
Intended use The Eurogentec MiRmaid miRNA precursors qRT-PCR primer set – designed by DNA Vision - was developed and optimized for the amplification of the human precursor microRNAs (pre-miRNAs) by performing a reverse transcription (RT) followed by a Real-Time qPCR using SYBR ® Green I assay. This product is sold for research use only. The kit is not intended for use in diagnostic or therapeutic procedures. Do not use internally or externally in humans or animals. All miRNAs that can be amplified with the MiRmaid miRNA precursors qRT-PCR primer set are listed in appendix 1 and 2 (as miRNA names evolve quickly along time you will find a conversion table on our website giving the new miRNA names corresponding to our miRNA MiRmaid names). Reference center: Dr. Ogier D. - INSERM U773 - Centre de recherche Bichat Beaujon - CRB3 - Paris - France Shipping and storage conditions The MiRmaid miRNA precursors qRT-PCR primer set kit is shipped on dry ice. Upon receipt, the MiRmaid RT primer MasterMIX, the 2 MiRmaid qPCR primer plates, the qPCR MasterMix Plus for SYBR ® Green I and the extra tube 50 mM MgCl 2 should be stored at -20 °C (-18 °C to –24 °C) in a constant temperature freezer to maintain stability. Repeated freeze-thaw cycles (more than 5) should be avoided. When stored under these conditions the reagents are stable for at least 1 year. The qPCR MasterMix Plus for SYBR ® Green I should be protected from light whenever possible. Safety information This product and its components should be handled only by persons trained in laboratory techniques and used in accordance with the principles of good laboratory practice. All chemicals should be considered potentially hazardous. Therefore, when handling chemical reagents, it is advisable that suitable protective clothing, gloves and eye/face protection are worn. Care should be taken to avoid contact with skin or eyes. In case of contact, wash immediately with water and seek medical advice. DMSO, in which SYBR ® Green I is dissolved, may cause eye, skin, and respiratory tract irritation. It is readily absorbed through the skin. DMSO is a combustible liquid and vapor that should be kept away from heat and flame. The product contains material, Manual for MiRmaid miRNA precursor qRT-PCR Primer Set I www.eurogentec.com I info@eurogentec.com 3
Page 1: MANUAL FOR MiRmaid miRNA PRECURSORS
Page 5 and 6: miRNA genes are usually transcribed
Page 7 and 8: MiRmaid miRNA precursors qRT-PCR pr
Page 9 and 10: 5S 18S 3. DNase and Reverse Transcr
Page 11 and 12: COMPONENT VOLUME PER WELL VOLUME FO
Page 13 and 14: Positive controls It is highly reco
Page 15 and 16: Troubleshooting guide This troubles
Page 17 and 18: The dissociation curves does not sh
Page 19 and 20: Homology between human pre-miRNA pr
Page 21 and 22: Ordering information Quantitative m
Page 23 and 24: Related products qPCR MasterMix plu
Page 25 and 26: Appendix 3: qPCR MasterMixes compat
Page 27 and 28: Worldwide Distribution INDIA GeneX

Safety information - Eurogentec

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