Removal of gDNA and synthesis of cDNA was performed with RNA products following the manufacturer’s instructions of ReverTra Ace qPCR RT Master Mix (Tiangen, Beijing, China). To validate the putative lncRNAs in V. destructor mites, 16 lncRNAs were randomly selected to determine with PCR amplification, which was carried out with the obtained cDNA in a 20 μL reaction volume mixture (2 × Taq PCR StarMix; GenStar, Beijing, China) on an Eppendorf cycler. PCR profile consists of a pre-denaturation at 94°C for 5 min; followed by 30 cycles including 94°C for 50 s, 55°C for 30 s, and 72°C for 1 min; and a final elongation step at 72°C for 10 min (Guo et al., 2018a (link)). The tested lncRNAs with their forward and reverse primers were presented in Supplementary Table 1 . PCR products were electrophoresed in 2.5% Tris acetate-EDTA-agarose gel containing 0.01% Gelview (BioTeke, Beijing, China) and visualized under ultraviolet light (Peiqing, Shanghai, China).
Revertra ace qpcr rt master mix
Manufactured by Tiangen Biotech
Sourced in China
ReverTra Ace qPCR RT Master Mix is a ready-to-use solution for reverse transcription and real-time PCR amplification in a single step. It contains all the necessary components for efficient cDNA synthesis and subsequent quantitative PCR.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using revertra ace qpcr rt master mix
1
Validating lncRNAs in Varroa Mites
2
Validating Varroa Destructor lncRNAs
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Synthesis of cDNA was performed with RNA products following the manufacturer's instructions of ReverTra Ace qPCR RT Master Mix (Tiangen, Beijing, China). To validate the putative lncRNAs in V. destructor mites, 16 lncRNAs were randomly selected to determine with PCR amplification, which was carried out with the obtained cDNA in a 20 μL reaction volume mixture (2×Taq PCR StarMix; GenStar, Beijing, China) on an Eppendorf cycler. PCR profile consists of a pre-denaturation at 94°C for 5 min; followed by 30 cycles including 94°C for 50 s, 55°C for 30 s and 72°C for 1 min; and a final elongation step at 72°C for 10 min [37] (link). The tested lncRNAs with their forward and reverse primers were presented in Table S1. PCR products were electrophoresed in 2.5% Tris acetate-EDTA-agarose gel containing 0.01% Gelview (BioTeke, Beijing, China) and visualized under ultraviolet light (Peiqing, Shanghai, China).
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