EvaGreen 2X qPCR MasterMix-ROX

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Application
Quantitative real-time PCR (qPCR)
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Purpose Guaranteed high-performance real-time PCR using EvaGreen 2X qPCR MasterMix.
Specificity QPCR Instruments:
- ABI® 7000, 7300, 7700, 7900, 7900HT,
- StepOnePlus™
- StepOne™
- OpenArray
- PRISM™ Sequencing Detection Series
- Biometra TOptical
- Fluidigm BioMarkTM
- Wafergene SmartChip System
- TianLong TL998 System
Characteristics - Enable streamlined protocol in a simple reaction set-up
- Allow accurate quantification of a variety of gene targets
- Reduce pipetting steps to minimize the risk of contamination
- Compatible with most real-time PCR instruments
Components Each EvaGreen 2X qPCR MasterMix is a 2X mixture of dNTPs, HotStart DNA Polymerase, Mg2+, fluorescent detection dye, reference dye and proprietary buffer components. Each set of EvaGreen qPCR MasterMix contains 4 vials, each containing 1.25 ml.
Application Notes - Gene-expression analysis
- Gene knockdown validation
- Microarray validation
- High throughput applications
- Virus detection and quantification
Comment

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
Storage -20 °C
Storage Comment Store at -20°C and protect from light. Store at 4°C if the MasterMix will be used within 2 weeks.
Product cited in: Kadam, Chuan: "Erratum to: Rectocutaneous fistula with transmigration of the suture: a rare delayed complication of vault fixation with the sacrospinous ligament." in: International urogynecology journal, Vol. 27, Issue 3, pp. 505, 2016 (PubMed). Method employed by authors: Quantitative real-time PCR (qPCR)

Su, Xu, Guan, Wang, Wu, Zhai, Chen, Hu: "Adjuvant effect of docetaxel on HPV16 L2E6E7 fusion protein vaccine in a mouse model." in: International immunopharmacology, Vol. 38, pp. 16-25, 2016 (PubMed).

Park, Ju, Park, Song, Shin, Lee, Jeong, Yu, Lee, Cho, Kim, Kim, Kim, Park, Chun: "PPARγ neddylation essential for adipogenesis is a potential target for treating obesity." in: Cell death and differentiation, Vol. 23, Issue 8, pp. 1296-311, 2016 (PubMed).

Wood, Cox, Phelps, Lai, Poddar, Talbot, Mu: "Thyroid Transcription Factor 1 Reprograms Angiogenic Activities of Secretome." in: Scientific reports, Vol. 6, pp. 19857, 2016 (PubMed).

Ren, Sun, Kang, Zhao, Feng, Ren, Han, Yang: "Responsiveness of soil nitrogen fractions and bacterial communities to afforestation in the Loess Hilly Region (LHR) of China." in: Scientific reports, Vol. 6, pp. 28469, 2016 (PubMed).

Zhao, Zhang, Guo, Li, Han, Song, Wang, Li, Li: "Identification and Validation of Reference Genes for qRT-PCR Studies of Gene Expression in Dioscorea opposita." in: BioMed research international, Vol. 2016, pp. 3089584, 2016 (PubMed).