Safety information - Eurogentec

More documents

Recommendations

Info

14 Typical results An assay performed with MiRmaid miRNA precursor qRT-PCR primer set is fast, sensitive and optimized to amplify the miRNA precursors. The following experimental data were obtained with 1 μg of total RNA extracted from lung cells. The assays were performed according to the procedure described above. Figure 4: Amplification plots using the qPCR MasterMix Plus for SYBR ® Green I (RT-SN2X-03+) and some miRNA primers from MiRmaid qPCR primer plates on an ABI Prism ® 7900 HT Figure 5: Melting-curve analysis of six miRNA precursor RT-qPCR products on the ABI Prism ® 7900 HT Manual for MiRmaid miRNA precursor qRT-PCR Primer Set I www.eurogentec.com I info@eurogentec.com
Troubleshooting guide This troubleshooting guide takes you through the most common difficulties you can meet when performing a SYBR ® assay with the MiRmaid miRNA precursors qRT-PCR primer set. Our scientific support is happy to help you if you might have any further questions (info@eurogentec.com / info.uk@eurogentec.com / info.usa@eurogentec.com). No product from experimental sample is amplified and positive control does not work as expected Forgot one of the component Check again if all the steps have been performed and verify the thermal cycler conditions. RNA has been damaged or degraded Replace RNA if necessary. Quality of RNA If RNA is not of good quality the DNase treatment can have a negative effect on the starting material > replace RNA if necessary. Check compatibility of RNA with this assay. Pay particular attention to the presence of small RNAs before and after DNase treatment (using positive control such as 5S). RNase contamination Maintain aseptic conditions and use RNase decontamination solution, add RNase inhibitor. Some miRNA precursors may not be expressed in your sample. MiRNA precursor expression profiles vary from one tissue to another or the expression of miRNA precursors may be too low to be detected. Increase the amount of starting material (to do so in step 16 of the protocol dilute the cDNA in a lower final volume). > We recommend performing positive control reactions using U6, 5S, TBP and HMBS in parallel with your sample. SYBR ® Green I has degraded SYBR ® Green I should be protected from light whenever possible. Manual for MiRmaid miRNA precursor qRT-PCR Primer Set I www.eurogentec.com I info@eurogentec.com 15
Page 1 and 2: MANUAL FOR MiRmaid miRNA PRECURSORS
Page 3 and 4: Intended use The Eurogentec MiRmaid
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: Positive controls It is highly reco
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

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?