Product category:
Polymerase chain reaction (PCR)
News Release from: Qiagen | Subject: QuantiTect
Edited by the Laboratorytalk Editorial
Team on 06 May 2005
Reverse transcription for two-step
RT-PCR
Genomic DNA removal is essential for accurate results in real-time RT-PCR assays that do not use RNA-specific primers
Reverse transcription for real-time, two-step RT-PCR is now faster and more convenient with the new QuantiTect reverse transcription kit, says Qiagen Only 20 minutes are required to synthesise first-strand cDNA and eliminate genomic DNA contamination
This article was originally published on Laboratorytalk on 20 Jun 2003 at 8.00am (UK)
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cDNA yields are high, allowing sensitive detection of even low-abundance genes in real-time, two-step RT-PCR.
Genomic DNA is effectively removed from starting RNA samples through a two-minute incubation in a unique buffer.
The treated RNA is then directly reverse transcribed without any further handling or incubation.
Genomic DNA removal is essential for accurate results in real-time RT-PCR assays that do not use RNA-specific primers.
Reverse transcription is conveniently set up using a master mix containing all reaction components, and then performed at the same temperature as the previous reaction for genomic DNA removal.
No separate steps are required for RNA denaturation, primer annealing, and RNase H digestion.
Quantiscript reverse transcriptase has a high affinity for RNA and is optimised for RNA amounts from 10pg to 1ug.
This high RNA affinity, in combination with Quantiscript RT buffer, enables high cDNA yields, even from templates with high GC-content or complex secondary structure.
RT Primer Mix contains an optimised mix of oligo-dT and random primers that enables cDNA synthesis from all regions of RNA transcripts, even from 5' regions.
This allows high yields of cDNA template for real-time PCR analysis regardless of where the target region is located on the transcript.
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