ReadyScript® cDNA Synthesis Mix

Protocol tips

Upstream tips	Protocol tips	Downstream tips
This Kit is optimized for the production of targets < 1kb in length. There will be no loss in functional performance after 20 cycles of freezing on dry ice and thawing on ice		For downstream reactions, use 1/5th to 1/10th of the first-strand reaction (2-4 μL) for PCR amplification

Upstream tips
This Kit is optimized for the production of targets < 1kb in length. There will be no loss in functional performance after 20 cycles of freezing on dry ice and thawing on ice
Downstream tips
For downstream reactions, use 1/5th to 1/10th of the first-strand reaction (2-4 μL) for PCR amplification

Publication protocol

Total RNA was isolated using RNeasy kit® (Qiagen, Hilden, Germany), and reverse transcribed in a thermal cycler using ReadyScript® cDNA Synthesis Mix (Sigma Aldrich). Amplification of cDNA was carried out in ViiA7 real-time PCR system with the following thermal profile (50 °C for 2 min and 95 °C for 10 min, then followed by 40 cycles of 95 °C for 15 s and 60 °C for 1 min). The TaqMan® Gene Expression of Cyp1a1 (Mm00487218_m1), AChE (Mm00477274_g1), Gapdh (Mm99999915_g1), and TaqMan® microRNA assay of has-miR-132 (ID:00457), has-miR-212 (ID:00515) and RNU6B (ID: 1093) were utilized in this study. System, kits and reagents for real-time PCR were purchased from Applied Biosystems (CA, USA). The relative gene expression was calculated using the comparative ∆∆Ct method.

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for ReadyScript® cDNA Synthesis Mix below.

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