QuantiTect Whole Transcriptome Kit (100)

QIAGEN offers this whole transcriptome amplification kit for unlimited real-time PCR analysis from limited RNA samples. The kit uses reverse transcription followed by ligation and multiple displacement amplification (MDA), integrating cDNA synthesis with REPLI-g technology for nonbiased sequence amplification. It is designed for whole total RNA amplification starting from purified total RNA, with a maximum input volume of 5 µl total RNA per reaction of 50 µl.

An optimized blend of reverse transcriptase and the highly processive Phi29 DNA polymerase enables uniform amplification of all transcript regions, including 5' ends, preserving the transcript profile for reliable gene expression analysis. Depending on amplification time, the procedure yields up to 10 µg cDNA in 2 hours or up to 40 µg cDNA in 8 hours, from as little as 1 ng purified total RNA (corresponding to roughly 50 cells) up to 300 ng without any change in transcript representation. RNA transcripts should be longer than 500 nucleotides, and the resulting amplified cDNA is approximately 20 kb in length, comprising concatemers of transcript sequences rather than full-length RNA transcripts.

The cDNA produced is intended for real-time PCR analysis using QuantiFast or Rotor-Gene Kits and can be safely archived for future studies. It is not suitable for microarray analysis or for amplification of short RNA species such as miRNA. The kit supports mid-throughput workflows with a minimal pipetting volume of 1 µl, and the generated cDNA does not require cleanup prior to real-time PCR.