The QuantiNova Reverse Transcriptase has a high affinity for RNA and is capable of cDNA synthesis from a wide range of RNA amounts (10 pg – 5 μg) (see figure “ Precise quantification from 5 µg – 10 pg RNA
”). The kit provides high yields of cDNA template for real-time PCR analysis, regardless of where the amplified target region is located on the transcript. Even difficult templates, such as those with high GC-content or complex secondary structure, are successfully reverse transcribed. The QuantiNova RT Buffer has also been optimized to be compatible with real-time PCR buffers.
To obtain accurate results in real-time RT-PCR gene expression assays, it is important that only cDNA is amplified and detected. Interference by genomic DNA can effectively and rapidly be removed using the gDNA Removal Mix (see figure “ Efficient removal of contaminating gDNA ensures precise quantification of transcripts
”). Time is saved and costs are reduced, since a separate DNase digestion during or after purification of RNA samples is not required. Also, designing RNA-specific primers or probes is not necessary.
Detecting variations in cDNA synthesis allows you to check the reproducibility of your results. The newly developed internal control is a defined transcript (RNA molecule) that can be optionally added to your samples and transcribed into cDNA. It is intended to report instrument or chemistry failures, errors in assay setup and the presence of inhibitors. Because the internal control behaves comparable to real transcripts, it can be used to confirm successful reverse transcription and amplification in subsequent qPCR (see figure “ Internal control reliably detects presence of inhibitors
Components of the QuantiNova Reverse Transcription Kit
|Component ||Benefits |
|gDNA Removal Mix ||Detection of RNA only in real-time RT-PCR |
|Internal Control RNA ||Verification of successful RT-PCR performance |
|QuantiNova Reverse Transcriptase ||Use of a wide range of RNA amounts (10 pg – 5 μg RNA) |
|QuantiNova RT buffer system ||Read-through of difficult templates |
|RT Primer Mix ||cDNA synthesis from all regions of transcripts, even from 5' regions |