iScript™ All-in-One RT Master Mix (5X)

iScript™ 5X All-In-One RT Master Mix is a convenient, ready-to-use formulation of all the reagents necessary for first-strand cDNA synthesis with the exception of the template. This optimized, 5X All-In-One RT Master Mix contains novel iScript™ IV Reverse Transcriptase (RT), RNase Inhibitor, dNTPs and a finely balanced ratio of Oligo(dT)₂₀ and Random Primers. Oligo(dT)₂₀ anneals selectively to the poly(A) tail of mRNAs. Random Primers do not require the presence of poly(A) and they are utilized for the transcription of mRNA 5’-end regions. The first-strand cDNA can be directly used as a template in PCR.

iScript™ IV Reverse Transcriptase (RT) is a novel recombinant reverse transcriptase that exhibits much higher efficiency in the first-strand cDNA synthesis from RNA templates with secondary structures and high GC content. The iScript™ IV Reverse Transcriptase is engineered to work under high temperatures (50-55°C), which can further facilitate to resolve the secondary structures and high GC problems of RNA. Besides, the iScript™ IV RT is significantly improved in inhibitor resistance, processivity, and reaction speed. iScript™ IV RT is designed to provide reliable, consistent, and fast cDNA synthesis in the presence of inhibitors found in a wide variety of samples.

Note: Upon completion of the first-strand cDNA synthesis, the cDNA product can be directly applied as a template in a standard PCR/qPCR.

Storage Conditions

Store all components at -20°C in a non-defrost cycle freezer. All components are stable for 1 year from the date of shipment when stored and handled properly.

Primer Information

Oligo(dT)₂₀ are oligonucleotides that anneal to the 3’-Poly(A) tail of mRNAs. Therefore, the utility of Oligo(dT)₂₀ is restricted to case scenarios where only mRNA or total RNA templates with 3’-Poly(A) tails are used for cDNA synthesis. On the other hand, since Random Primers anneal at non-specific sites within RNA template(s), they can be used generically for all forms of RNA as template for cDNA synthesis.