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IgScript™ Probe-Based One Step RT-qPCR 2x master mix contains IgScript™ Reverse Transcriptase, Taq DNA polymerase, MgCl2, dNTPs, stabilizers and low ROX reference dye with standard buffer providing improved RT-qPCR efficiency, wider dynamic range, superior sensitivity and specificity. IgScript™ Probe-Based One Step RT-qPCR Kit is a ready-to-use cocktail containing all components except primers, probe and template, for the amplification and detection of DNA in RT-qPCR. This 2x master mix requires minimal handling during reaction setup and offer consistent and robust RT-qPCR reactions.
IgScript™ Reverse Transcriptase is a recombinant MMLV reverse transcriptase with reduced RNase H activity, increased thermostability and can produce cDNA from small amount of total RNA for real-time RT-qPCR analysis and other applications. Taq DNA Polymerase is a thermostable DNA polymerase that possesses a 5´→3´ polymerase (1, 2) and a 5´→3´ exonuclease activity (3, 4). The amplification step features a high quality Taq DNA Polymerase which offers robust, reliable and better amplification.
Probe-Based One Step RT-qPCR Kit Includes:
IgScript one step RT-qPCR 2x master mix
Applications
Gene expression data validation.
Multiplexing
Mutation detection
Pathogen and viral detection
Genetically modified organisms (GMO) characterization and Genetic profiling
Benefits
Enhanced efficiency, specificity, and sensitivity
Compatible with all real-time PCR instruments
Superior gene expression results under various cycling conditions
Robust and active for cDNA synthesis at temperatures up to 55°C.
Mix and then centrifuge briefly to collect contents at the bottom of the tube.
Prepare a master mix for each reaction and control requiring Reverse Transcriptase enzyme plus 10% extra to allow for pipetting error according to the following table:
Mix the reaction mixture thoroughly.
Program the thermal cycler according to the manufacturer’s instructions.
A typical PCR cycling program is outlined in the following table.
Place the PCR tubes in the thermal cycler and start the cycling program.
Analyze the data according to manufacturer protocol.
* For 3 step cycling protocols, anneal at optimal annealing temperature for 30 sec followed by the minimum time required for data acquisition at 72 ºC according to instrument guidelines.
