EvaGreen miRNA qPCR MasterMix-iCycler

Details for Product No. ABIN4219201,
Quantitative real-time PCR (qPCR)
Purpose EvaGreen miRNA qPCR MasterMix is designed specifically for quantitative real-time analysis of miRNA samples.
Specificity The components of EvaGreen miRNA qPCR MasterMix have been developed for superb performance in sensitivity, signal-to-noise ratio, and complete elimination of primer dimers. The chemically modified Hotstart Taq polymerase included in our MasterMix significantly reduces non-specific PCR amplification observed with regular Taq polymerase.
Characteristics Primer concentration should not be high. 100nM to 300nM each
- primer usually gives best results
- Melting curve is useful in distinguishing nonspecific amplification products and primer dimers from desired product. It's recommended to run melting curve following amplification cycles.
- To do replicates
- To reduce pipetting error: preparing amplicon specific master mix, using repeating pipet, and keeping pipetting volume in manufacture suggested range are very effective to get tight Ct.
Application Notes - Gene-expression analysis
- Gene knockdown validation
- Microarray validation
- High throughput applications
- Virus detection and quantification

4 X 1.25 ml, for 500 reactions (20 μl)
1. Aliquot reagents to avoid contamination and to avoid repeated freeze-thaw cycles.
2. Primer concentration should not be high, a concentration of 100 nM to 300 nM of each primer usually gives the best results.
3. A very effective way to get tight Ct among replicates is to reduce pipetting error, this can be achieved by: preparing amplicon specific pre-mix, using repeating pipet, and keeping pipetting volume in manufacture suggested range.
4. For optimal results, it is recommended that the primers are 18-22 nucleotides in length with a Tm of 58°C-60°C and the size of target is about 100-250 bp.

Restrictions For Research Use only
Concentration 2 x
Storage -20 °C
Storage Comment Upon arrival, EvaGreen miRNA qPCR MasterMix should be stored at -20°C and protected from light. After each experiment, the leftover thawed mix can be stored at 4°C if it is to be used within the next 3 months.