EvoTM RT Mastermix (with cell lysis) is a 5X All-In-One RT Mastermix (with Cell Lysis Kit) that offers a quick, simple and robust method to prepare template for first-strand cDNA synthesis directly from cultured cells, enabling reverse transcription of lysates from 10-105 cultured cells without time-consuming and hazardous-chemicals-involved RNA extraction and purification steps. The kit includes reagents for cell lysis as well as genomic DNA (gDNA) removal. The presence of contaminating gDNA in RNA preparations is often a significant problem for downstream applications, leading to false-positive signals and misinterpretation of gene expression levels. The lysis procedure simultaneously eliminates genomic DNA effectively in 12 min, without compromising RNA quality, and therefore ensures consistent, reproducible, and accurate results with 10-105 cells.
The lysate can then be directly reverse-transcribed into cDNA using the 5X all-in-one RT Mastermix. The 5X All-In-One RT Mastermix is a ready-to-use formulation of all the reagents necessary for first-strand cDNA synthesis, including proprietary Evo™ Reverse Transcriptase (Evo™ RTase), RNaseOFF Ribonuclease Inhibitor, dNTPs and a finely balanced ratio of Oligo(dT)s and Random Primers. Coupled together, this complete system provides the ultimate convenience in generating high-quality cDNA directly from 10-105 cultured cells, suitable for a wide range of downstream applications. Note: Upon completion of the first-strand cDNA synthesis, the cDNA product can be directly applied as a template in a standard PCR/QPCR.