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.
|2X miRNA cDNA Synthesis SuperMix||20 rxn (200 μl)|
|Enzyme Mix||20 rxn (40 μl)|
|Nuclease-Free H2O||1.0 ml|
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
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.