G/G (wild type)
The reverse transcription reaction is the first critical step of RT-PCR procedures. To ensure that this reaction is optimized, Stratagene has included StrataScript Rnase H-minus RT in both the single-step and two-step formats of the Brilliant and Brilliant Plus QRT-PCR core reagent kits. StrataScript reverse transcriptase is a genetically modified Moloney murine leukemia virus reverse transcriptase with a point mutation that results in removal of RNase H activity. In numerous tests, StrataScript reverse transcriptase has produced high yields of cDNA and long transcripts, providing ideal cDNA templates for subsequent PCR amplifications.4
The Brilliant single-step QRT-PCR core reagent kits include all the necessary reagents for cDNA synthesis and PCR amplification in a single tube. This format uses primers specific to the RNA of interest. Although UNG cannot be used in the single-step format, the Brilliant Plus kit includes dUTP in the nucleotide mix. Should any amplicon be released from the single-step RT-PCR, the decontamination control would then eliminate amplification of contaminating DNA in subsequent PCR or two-step RT-PCR.
The Brilliant two-step QRT-PCR core reagent kit is ideal for use when
sensitivity of the reverse transcription reaction needs to be increased. This
increased sensitivity is achieved through the provided buffers that are
specifically optimized for the first-strand synthesis and for the P