Storage Condition
-20℃
Description
All-in-One First-Strand Synthesis MasterMix is a high-quality, efficient, and convenient one-step cDNA synthesis kit. It is designed to minimize contamination and contains all the necessary components for first-strand cDNA synthesis, including thermostable M-MLV GIII Reverse Transcriptase and its reaction buffer, RNase inhibitor, dNTPs, Oligo(dT)20VN, and random primers-all the necessary components. Simply add RNA template and water to initiate the reaction. The cDNA obtained using this kit is primarily used for downstream qPCR experiments.
Protocol
① Prepare the following reaction mixture on ice:
② Mix gently and spin down.
③ Incubate the mixture at 50℃ for 15 minutes.
Note: If the template RNA does not contain a poly(A) tail, you may pre-incubate 25℃ for 10 minutes.
④ Terminate the reaction by incubating at 85℃ for 5 minutes.
⑤ Place the obtained cDNA solution on ice for subsequent experiments.
Note: The cDNA solution can be stored at -20℃ for up to one week. Long-term storage is recommended at -80℃ .
Notice
The premix contains Oligo(dT)20VN and random primers, suitable not only for eukaryotic mRNA containing poly(A) tail, but also for templates that do not contain poly(A) tail, such as prokaryotic RNA, eukaryotic rRNA, tRNA, etc. However, it is not suitable for small RNA like miRNA.
Since random primers can initiate reverse transcription at any position on the RNA, it is not recommended to use this product for fulllength cDNA cloning of eukaryotic organisms. For obtaining full-length cDNA of eukaryotic organisms, we recommend RTase III Primer Flexible All-in-One Mix .