igScript™ One Step RT-qPCR Kit

$195.00 – $695.00

Description

igScript™ One Step RT-qPCR kit combines two powerful mixtures: i). igScript™ Reverse Transcriptase and ii) ig SYBR® Green qPCR 2x master mix with standard buffer providing improved PCR efficiency, wider dynamic range, superior sensitivity and specificity. The two mixtures require 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 and increased thermostability. The kit is highly efficient at producing full-length cDNA from long RNA templates at temperatures between 42-55ºC.

Intact Genomics SYBR® Green qPCR 2x master mix is a ready-to-use cocktail containing all components except primers and template, for the amplification and detection of DNA in qPCR. The Ig SYBR® Green qPCR 2x master mix with integrated chemically-modified hot start Taq DNA polymerase, SYBR® Green I fluorescent dye, ROX dye, MgCl2, dNTPs and stabilizers. This master mix is ideal for high-throughput real-time PCR screening and validation. The amplification step features a high quality hot start Taq DNA Polymerase which offers higher fidelity and better amplification.

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.

igScript™ One Step RT-qPCR Kit Includes

ig Script™ Reverse Transcriptase
ig SYBR™ Green qPCR 2x Master Mix

Storage Temperature
–20 °C4214 4218

Additional information

reaction	100 reactions (20 µl rxn vol), 500 reactions (20 µl rxn vol)

Protocol

Place kit components and RNA samples on ice.
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.