Accuris qMax™ Green Reverse Transcription quantitative PCR kit (RT-qPCR): Reverse-transcribes RNA into complementary DNA (cDNA), then uses cDNA as starting template for amplification of gene target of interest by means of qPCR.
Two-step reaction (i.e. Reverse Transcription plus DNA Amplification) is performed in one reaction tube with a single reaction setup.
Amplified DNA is detected by means of an intercalating (i.e. dsDNA-binding) fluorescent green dye, qMax Green, which is similar to SYBR Green, but exhibits higher fluorescence and less PCR inhibition.
qPCR, also known as Real-Time PCR, allows the user to amplify DNA by means of PCR and monitor the progression of amplification quantitatively, in real time. That is, qPCR allows the user to measure quantities of amplified target DNA, cycle by cycle.
qMax Green One Step RT-qPCR Kit contains the following components:
- 2X Master Mix (containing Accuris Hot Start Taq Polymerase, dNTPs, qMax Green dye, buffer that includes PCR enhancers and inert blue dye that visually aids the user during pipetting)
- 20X Reverse Transcriptase (an RNA-dependant DNA polymerase, derived from MMLV, and an RNase Inhibitor)
→Available with low or high levels of ROX (select in dropdown menu). ROX is a fluorescent, passive reference dye that is used to normalize the signal emitted from the reporter dye, qMax Green, during qPCR.
The fluorescent signal from the passive dye remains constant during qPCR, while the signal from the reporter dye changes as cycles progress. Real-Time PCR instruments have varying requirements for ROX levels.
Kit is compatible with standard and fast-cycling PCR protocols.
→Available in 10 (sample size), 100, 500 and 1,000 reactions sizes (select in dropdown menu). Reaction sizes are based on total reaction volumes of 20 uL.
Accuris catalog numbers:
Low ROX: 10 rxns (PR2120-L-S), 100 rxns (PR2120-L-100), 500 rxns (PR2120-L-500), 1,000 rxns (PR2120-L-1000)
High ROX: 10 rxns (PR2120-H-S), 100 rxns (PR2120-H-100), 500 rxns (PR2120-H-500), 1,000 rxns (PR2120-H-1000)
The following link to ThermoFisher Scientific provides a good understanding and overview of qPCR: Essentials of Real-Time PCR