miRNA All-In-One cDNA Synthesis Kit
miRNA All-In-One cDNA Synthesis Kit is a complete system for efficient synthesis of first strand miRNA from total RNA templates. This comprehensive kit comes with just two components: 2X miRNA cDNA Synthesis SuperMix and Enzyme Mix, enabling hassle-free set-ups with minimal potential for contamination due to significantly reduced pipetting. The kit contains all miRNA synthesis-required reagents including Poly(A) Polymerase and abm’s OneScript® Plus Reverse Transcriptase, a unique miRNA Oligo d(T) adapter that anneals to the A-tailed miRNA template, and RNaseOFF Ribonuclease Inhibitor to protect RNA template from degradation.
Store at -20°C.
|Unit quantity||20 rxn|
|Do I need to purify the sample after adding the poly A tail from the Ambion kit before using the cDNA synthesis kit?|
No, that is not necessary. As per page 4 in the miRNA manual http://www.abmgood.com/miRNA/pdfs/miRNA%20HandbookV11.pdf, the sample can be used directly for the next step in the cDNA synthesis kit.
|How to design individual miRNA primer ?|
The individual miRNA primer could be purchased at http://www.abmgood.com/miRNA-Primer-Sets-Human.html
|Is it possible to use your kit (miRNA cDNA Synthesis Kit) to do quantitative PCR with the Taqman strategy?|
This kit along with our miRNA primers would not be compatible for use with Taqman strategy.
|How does the universal reverse primer work?|
The universal reverse primer can be used for all mature miRNA. To quantify or amplify a specific miRNA, the universal reverse primer with the appropriate forward miRNA primer needs to be used. Our forward primers can work with any kits. Our universal primers will only work with abm's cDNA synthesis kit.
|Can I use SYBR green mastermix for downstream qPCR of the cDNA produced with this kit?|
It may be possible to use a SYBR green mastermix for qPCR, however, our BrightGreen miRNA miRNA qPCR MasterMix recipe has been fully optimized to provide the most successful conditions for qPCR. We have performed extensive testing with minor differences in the composition of miRNA qPCR buffer, this has proven to give very dramatic differences to the results of qPCR. General qPCR mastermixes have more additives to prevent non-specificity and formation of primer dimers and the conditions are often too harsh for the miRNA primers to anneal properly
|Should DNase treatment be performed for the RNA extracted before this procedure?|
You can perform gDNA removal before proceeding with the cDNA synthesis. However, we would recommend using Cat# G488 AccuRT gDNA removal kit. Our product uses proprietary method to removal gDNA and to stop the gDNA removal without using chelating agent such as EDTA and no heating step is involved.
|Can the miRNA cDNA Synthesis Kits also be used for plant miRNAs?|
Yes, these kits can also be used for plant miRNAs.
|Can this kit be used for circulating miRNA or exosomal miRNA?|
Yes, our kit does work with circulating miRNA and exosomal miRNA. Depending on the abundance of the target gene, you may use a lower template amount. E.g. House-keeping genes starting template of 100ng total RNA may be sufficient.
- Lee, G et al. "Potential monoclonal antibody therapy for the treatment of ovarian cancer" Ovarian cancer—basic science perspective :385-406 (2012). DOI: 10.5772/1268 . Application: Reverse Transcription.
- Ramraj, et al. "Serum-circulating miRNAs predict neuroblastoma progression in mouse model of high-risk metastatic disease" Oncotarget 7.14:18605 (2016). DOI: 10.18632/oncotarget.7615. Application: miRNA qPCR Analysis.
- Duan, X. L., Cong-Cong, M., Hua, J., Xiao, T. W., & Luan, J. "Benzyl butyl phthalate (BBP) triggers the malignancy of acute myeloid leukemia cells via upregulation of PDK4" Toxicology in Vitro 104693: (2019).
- Gomaa, A., Peng, D., Chen, Z., Soutto, M., Abouelezz, K., Corvalan, A., & El-Rifai, W. "Epigenetic regulation of AURKA by miR-4715-3p in upper gastrointestinal cancers" Scientific reports 9(1):1-11 (2019).
- Liang, Y., Song, X., Li, Y., Su, P., Han, D., Ma, T., … Yang, Q. "circKDM4C suppresses tumor progression and attenuates doxorubicin resistance by regulating miR-548p/PBLD axis in breast cancer" Oncogene 38(42):6850–6866 (2019). DOI: 10.1038/s41388-019-0926-z.
- Verma, M., Asakura, Y., & Asakura, A. "Inhibition of microRNA‐92a increases blood vessels and satellite cells in skeletal muscle but does not improve duchenne muscular dystrophy–related phenotype in mdx mice" Muscle & Nerve 59(5):594–602 (2019). DOI: 10.1002/mus.26433.
- Wu., Chien-Wei., . "Downregulation of MiR-144 by Triptolide Enhanced p85α−PTEN Complex Formation Causing S Phase Arrest of Human Nasopharyngeal Carcinoma Cells" European Journal of Pharmacology 855:137-148 (2019). DOI: 10.1016/j.ejphar.2019.04.052..
- Wu, F., Niu, Z., Zhou, B., Li, P., & Qian, F. "PSMB1 Negatively Regulates the Innate Antiviral Immunity by Facilitating Degradation of IKK-ε" Viruses 11(2):99 (2019). DOI: 10.3390/v11020099.
- Wu, M.-J., Chen, Y.-S., Kim, M. R., Chang, C.-C., Gampala, S., Zhang, Y., … Chang, C.-J. "Epithelial-Mesenchymal Transition Directs Stem Cell Polarity via Regulation of Mitofusin" Cell Metabolism 29(4):993–1002.e6 (2019). DOI: 10.1016/j.cmet.2018.11.004.
- Xiang, X., Zhou, Y., Sun, H., Tan, S., Lu, Z., Huang, L., & Wang, W. "Ivabradine abrogates TNF-α-induced degradation of articular cartilage matrix" International Immunopharmacology 66:347–353 (2019). DOI: 10.1016/j.intimp.2018.11.035.
- Zhao, G., Zhang, T., Sun, H. J., & Liu, J. "Copper nanoparticles induce zebrafish intestinal defects via endoplasmic reticulum and oxidative stress" Metallomics. : (2019).
- Zhou, L., Li, P., Zhang, M., Han, B., Chu, C., Su, X., ... & Ma, S. "Carbon black nanoparticles induce pulmonary fibrosis through NLRP3 inflammasome pathway modulated by miR-96 targeted FOXO3a" Chemosphere 125075: (2019).
- Zhou, Y., Lei, J., Xie, Q., Wu, L., Jin, S., Guo, B., ... & Zhang, J. "Fibrinogen-like protein 2 controls sepsis catabasis by interacting with resolvin Dp5" Science Advances 5(11):eaax0629 (2019).
- Zhu, R., Xue, X., Shen, M., Tsai, Y., Keng, P. C., Chen, Y., … Chen, Y. "NFκB and TNFα as individual key molecules associated with the cisplatin-resistance and radioresistance of lung cancer" Experimental Cell Research 374(1):181–188 (2019). DOI: 10.1016/j.yexcr.2018.11.022.