All Script RT Master Mix is a Real Time PCR reverse transcription reaction kit that adds the function of removing genomic DNA (gDNA), using our company's new high-efficiency reverse transcriptase. The total RNA purified by common RNA extraction methods often contains trace amounts of gDNA. When reverse transcribing the cDNA obtained through Real Time PCR for gene expression analysis, if there are pseudogenes detected in the target gene or primers cannot be designed across introns, the mixed gDNA will be used as a template for amplification, affecting the accuracy of amplification results. This product is pre mixed with a thermosensitive DNase with strong DNA decomposition activity. This DNase is effective, efficient, and easily inactivated. With just one step, cDNA without gDNA can be easily obtained.
The cDNA synthesized using this product is suitable for qPCR analysis using both dye and probe methods. According to the experimental purpose, quantitative PCR reagents such as Universal Super SYBR Master Mix (PCM60) can be used in combination.
Feature
Can easily and quickly remove genomic DNA and synthesize cDNA All gDNA removal reaction reagents and reverse transcription reaction reagents have been pre mixed, and gDNA removal and cDNA synthesis can be achieved in about 15 minutes through a single tube reaction.
High thermal stability Perform reverse transcription reaction at 50 ℃.
Can perform uniform reverse transcription reactions on the entire region of RNA The reaction buffer most suitable for cDNA synthesis in RealTime PCR and Primermix (Oligo dT and Random Primer) mixed in the optimal ratio were used to achieve uniform and efficient reverse transcription of the entire region of RNA.
High adaptability to Real Time PCR reagents The components that have the least impact on the reaction system of Real Time PCR were used, and even when a maximum of 20% reverse transcription reaction solution was introduced into the PCR reaction solution, it still showed good linearity. Therefore, this kit is suitable for the detection of low abundance mRNA.
Note
This product uses a thermosensitive DNase, please make sure to place the 5 x All Script RT Master Mix on ice.
Before using the 5×All Script RT Master Mix, please briefly centrifuge and collect it to the bottom of the tube, and gently blow with a pipette to mix thoroughly before accurately aspirating.
During the operation, RNase contamination should be avoided. RNase Free Water used for reverse transcription and PCR reactions should be stored separately from RNase Free Water used for other experiments to avoid sharing.