The Luna SARS-CoV-2 RT-qPCR Multiplex Assay Kit contains all the necessary components to conduct multiplex RT-qPCR assays as specified in the details below.
- The Luna Probe One-Step RT-qPCR 4X Master Mix with UDG (NEB #M3019) is supplied at 4X concentration and contains all the necessary components for One-Step RT-qPCR. It is also formulated with a unique passive reference dye that is compatible across a variety of instrument platforms, dUTP/UDG for carryover prevention and a nonfluorescent visible dye for monitoring reaction setup.
- The SARS-CoV-2 Primer/Probe mix provided in this kit contains primers and probes specific to two regions of the SARS-CoV-2 virus N gene [based on sequences provided by the Centers for Disease Control and Prevention (CDC)]. However, they have been modified to contain different fluorophores (N1, HEX; N2, FAM) to enable simultaneous observation on two different channels of a real-time instrument. To ensure the integrity of the input material and absence of inhibition, an internal control (IC) primer and probe set, designed to amplify the human RNaseP gene, is also provided in the primer mix. The reverse primer of this target has been modified from the CDC design to target an exon/exon boundary to reduce background amplification from possible contaminating genomic DNA. Amplification of the IC is observed in the Cy5 channel.
- A positive control (PC) template (SARS-CoV-2 N gene cloned into a plasmid) is also provided.