FAQ-111
Is mRNA isolation necessary for sensitive RT-PCR?
Usually not. Since RT-PCR is extremely sensitive, as little as 10–200 ng total RNA is sufficient for each 25–50 µl reaction mix, depending on the RT system. For abundant mRNA species, it is possible to use even less than 10 ng total RNA. For rare mRNA species, use a sequence-specific primer in the RT reaction to increase sensitivity. RNA content in various cells and tissues can be found here.
Related products
TurboCapture mRNA Kits
The TurboCapture System uses unique oligonucleotide immobilization technology to provide a fast and simple procedure for high-throughput mRNA extraction. Cell lysates are added to the wells of a TurboCapture plate (96 or 384 wells), and mRNA is allowed to hybridize to the immobilized oligo-dT in each well. Contaminants are washed away, and the isolated mRNA is then either used directly in cDNA synthesis or eluted for use in downstream applications.
QuantiTect Reverse Transcription Kit
The unique QuantiTect Reverse Transcription Kit provides a fast and convenient procedure for cDNA synthesis with integrated genomic DNA removal. Genomic DNA contamination in RNA samples is effectively eliminated by gDNA Wipeout Buffer. All components that are required for fast and efficient reverse transcription are provided with the QuantiTect Reverse Transcription Kit, including Quantiscript Reverse Transcriptase, Quantiscript RT Buffer, and a unique RT Primer Mix. The synthesized cDNA is optimized for use in real-time PCR, allowing reliable quantification of targets from all regions of mRNA transcripts.
Sensiscript RT Kit
The Sensiscript Reverse Transcriptase (RT) Kit is ideally suited for highly sensitive applications using very small amounts of RNA (less than 50 ng), such as single-cell RT-PCR and analysis of biopsies and LMD samples. Sensiscript Reverse Transcriptase allows highly efficient RT-PCR over a wide dynamic range and extremely sensitive RT-PCR with very small RNA amounts due to its high affinity for RNA. The Sensiscript RT Kit includes Sensiscript Reverse Transcriptase, dNTPs, and an optimized reaction buffer. Simply add primers for fast and easy cDNA synthesis.
QIAGEN OneStep RT-PCR Kit
The QIAGEN One-Step RT-PCR Kit provides a blend of Sensiscript and Omniscript Reverse Transcriptases, HotStarTaq DNA Polymerase, QIAGEN OneStep RT-PCR Buffer, a dNTP mix, and Q-Solution, a novel additive that enables efficient amplification of "difficult" (e.g., GC-rich) templates. The easy one-tube setup and optimized components result in highly sensitive and successful results.
Omniscript RT Kit
The Omniscript Reverse Transcription (RT) Kit is specially designed for reverse transcription with any amount of RNA from 50 ng to 2 μg per reaction. A high affinity for RNA allows Omniscript Reverse Transcriptase to provide superior performance compared with other reverse transcriptases, delivering higher sensitivity in RT-PCR, even with low-copy numbers. The Omniscript RT Kit includes Omniscript Reverse Transcriptase, dNTPs and an optimized reaction buffer. Simply add primers for fast and easy cDNA synthesis.