| Countries |

Research Diagnostic Therapeutic

The Eurogentec MiRmaid miRNA precursors qRT-PCR primer set (human) – 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.

Name	Quantity	Reference	EUR
MiRmaid miRNA precursors qRT-PCR primer set (human) + qPCR MasterMix Plus for SYBR® Green I, 7.5 ml	RT primers for 50 RTF&R qPCR primers for 25 qPCR reactionsqPCR MasterMix SYBR® for 600 qPCR reactions	RT-SYMI-025	1650.00
MiRmaid miRNA precursors qRT-PCR primer set (human) + qPCR MasterMix Plus for SYBR® Green I No ROX	RT primers for 50 RTF&R qPCR primers for 25 qPCR reactionsqPCR MasterMix SYBR® No ROX for 600 qPCR reactions	RT-SYMI-025NR	1650.00
MiRmaid miRNA precursors qRT-PCR primer set (human)	RT primers for 50 RTF&R qPCR primers for 25 qPCR reactions (25 µl)	RT-OLMI-025SET	1350.00

25 µl qPCR reactions

Delivery times

2 WD if ordered before Thursday

Packaging

The MiRmaid miRNA precursors qRT-PCR primer set contains:
- RT primer Master MIX (red cap): one tube of specific miRNA RT primers at 10 µM each. Sufficient to perform 50 RT reactions (25 samples)
- 170 qPCR primer sets containing 112 µl of the specific miRNA Reverse and Forward primers in each well.
- 4 positive qPCR control sets containing 112 µl of the specific miRNA Reverse and Forward primers in each well
These 170 pre-miRNA F and R primer mixes and the 4 positive control mixes are sufficient to performed at least 25 Real-Time qPCR reactions (25 µl reactions).
- 1 qPCR MasterMix Plus for SYBR® Green I, 7.5 ml, sufficient to performed at 600 Real-Time qPCR reactions (25 µl reactions).

Shipping conditions

Dry ice

Storage conditions

The MiRmaid miRNA precursors qRT-PCR primer set kit is shipped on dry ice.
Upon receipt, the MiRmaid RT primer Master MIX, the 2 MiRmaid qPCR primer plates, the qPCR MasterMix Plus for SYBR® Green I and the extra tube 50mM MgCl2 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.
qPCR MasterMix Plus for SYBR® Green I should be protected from light whenever possible.

The Eurogentec MiRmaid miRNA precursors qRT-PCR primer set
- designed by DNA Vision - was developed and optimized for the amplification of the human precursor miRNAs (pre-miRNAs) by performing a reverse transcription (RT) followed by a Real-Time qPCR using SYBR® Green I assay.
The Eurogentec MiRmaid miRNA precursors qRT-PCR primer set contains sufficient MiRmaid RT primer MasterMix to perform 50 RT (Reverse Transcriptase) reactions, which is enough for 25 different samples. The amount of cDNA produced after 1 RT reaction should allow performing 96 qPCR reactions.

The 2 MiRmaid qPCR primer plates contain specific pre-miRNA Reverse and Forward primers in each tube. These 170 pre-miRNA Forward and Reverse primer mixes and the 4 positive control mixes are sufficient to performed at least 25 Real-Time qPCR reactions.
Four different controls have been included in the 2 MiRmaid qPCR primer plates. U6 and 5S are small size RNAs comparable to the size of pre-miRNAs. By amplifying the U6 and 5S positive controls by Real-Time qPCR you will be able to check that the qPCR reaction have been performed correctly and that any small RNAs have been well extracted, reverse transcribed to cDNA and then have been amplified correctly.
Pre-miRNAs are very low expressed RNAs, however 6U and 5S positive controls are highly expressed RNAs, this is why 2 others low expressed positive controls, HMBS and TBP, have been added. These 2 positive controls are coming out at Ct values that are more or less comparable to the Ct values expected when amplifying precursor miRNAs.

The MiRmaid miRNA precursors qRT-PCR Primer Set (Human) has been designed and validated on ABI Real-Time thermocyclers and on DNA Engine Opticon® / Chromo 4 (Biorad) Real-Time thermocyclers. However the MiRmaid miRNA precursor qRT-PCR Primer Set (Human) could also be used on other platforms.
Eurogentec can provide you with the correct kit according to your Real-Time thermocycler. Please refer to the compatibility table relating each of our qPCR MasterMix for SYBR® Green I for the use on each Real-Time thermocyclers.

List of the precursor miRNAs present in the MiRmaid miRNA precursors qRT-PCR Primer Set (human)

Please download the list of the 170 pre-miRNAs that can be amplified and quantified with the MiRmaid miRNA precursors. This excel file also presents the human pre-miRNA primer sets that could be used for the amplification of the mouse or rat pre-miRNAs. The MiRmaid miRNA precursors qRT-PCR Primer Set is also available as MiRmaid miRNA precursors qRT-PCR individual sets. Please follow the following link to access to the list of reference code for each MiRmaid miRNA precursors qRT-PCR individual set available.

Standard protocol

Typical results

An assay performed with the MiRmaid miRNA precursor qRT-PCR primer set is fast, sensitive and optimized to amplify the pre-miRNA.
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.
Reference Center: Dr Ogier D., INSERM U773 - Centre de recherche Bichat Beaujon - CRB3 - Paris - France.

MiRmaid typical results
Amplification plots and meltcurve analysis using the qPCR MasterMix Plus for SYBR® Green I and some miRNA primers from the MiRmaid miRNA precursor qRT-PCR primer set on an ABI prism 7900HT®.

Legal notices

FOR RESEARCH USE ONLY - NOTICE TO PURCHASER: LIMITED LICENSE - Licensed dsDNA-Binding Dye Research Kits - Use of this product is covered by one or more of the following US patents and corresponding patent claims outside the US: 5,079,352, 5,789,224, 5,618,711, 6,127,155, 5,677,152, 5,773,258, 5,407,800, 5,322,770, 5,310,652, 5,994,056, 6,171,785, and claims outside the US corresponding to US Patent No. 4,889,818. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser’s own internal research. No right under any other patent claim (such as apparatus or system claims in US Patent No. 6,814,934) and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser’s activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patents require a separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA. Cyclic-substituted unsymmetrical cyanine dyes are covered by U.S. Patents 5,436,134 and 5,658,751 and licensed to Eurogentec S.A by Molecular Probes, Inc in the direct research field.

Cyclic-substituted unsymmetrical cyanine dyes are covered by U.S. Patents 5,436,134 and 5,658,751 and licensed to Eurogentec S.A by Molecular Probes, Inc in the direct research field.

Use of UDG employs U.S. Patents 5,035,996, 5,945,313, 5,683,896 and their foreign counterparts licensed to Eurogentec, S.A. from Invitrogen Corporation.

Manual

Leaflets

MSDS

Certificate of analysis

To obtain the Certificate of Analysis corresponding to your qPCR kit of interest please contact us via email and display the reference code of the qPCR kit and the batch number. Contact us

How to order by e-mail

Send your orders by email. To get the template ordering form, please contact our Customer Help Desk or send an e-mail to info@eurogentec.com.
Do not hesitate to contact us to set up the best ordering method for you.

How to order by fax

It is possible to order your product by fax; please check the fax numbers at the bottom of each page of our catalogue or below. This ordering method requires more manpower than the previously mentioned ones any specifications in our system. Moreover we are applying a double check to avoid any mistakes. The treatment of these orders requires one additional working day to the standard delivery times. For fax orders an additional fee of 2.50 eur per product will be charged. However, we do not recommend fax ordering, because these orders have to be processed manually and are therefore errorprone.

How to order by mail

It is possible to order your MirMaid products by mail; please use the following adress:

Do not hesitate to contact us for any additional information or comments on these ordering tools.