What is the maximum cDNA length that can be synthesized by QuantiTect Reverse Transcription Kit?
QuantiTect Reverse Transcription kit is specifically developed for real-time PCR. Amplicons for real-time are usually within the range of 70-200 bp and hardly exceeds 300 bp.
QIAGEN has tested this kit with amplicons up to 1.8 kbp. In this case, we suggest two modifications of the protocol.
- For amplicons > 500 bp, extend the reverse transcription time (30 minutes instead of 15 minutes)
- For amplicons > 1000 bp, use either oligo-dT only or a gene-specific primer. Combination of random primers + oligo-dT (as provided in kit) is not optimal in this case.