QuantiTect Reverse Transcription Kit From Qiagen

Both real-time polymerase chain reaction (PCR) and reverse transcriptase (RT) - PCR are common techniques used in almost every molecular laboratory. These techniques are very sensitive and allow the amplification and quantification of specific nucleotide sequences across several orders of magnitude. The usual starting point for both of these techniques is the extraction of high quality RNA from tissues or cells of interest. This RNA can then be used as a template to produce cDNA. Qiagen’s Quantitect Reverse Transcription Kit is fast and effective and will maximize cDNA yields when starting the reaction with high-quality RNA.

Not only does this kit provide high cDNA yields, it includes a step to eliminate any genomic DNA contamination. Additionally, a mastermix, containing reverse transcriptase, buffer and primers, is provided with the kit, which simplifies the procedure for production of cDNA. Other methods for generation of cDNA can involve making a mastermix from the purchase of separate components, which can lead to errors and contamination of the desired product. In addition, little starting material is required with the ability to produce cDNA from just picograms of RNA. Reverse transcriptase is the key requirement for cDNA synthesis and has the following three activities: an exoribonuclease, DNA-dependent DNA polymerase and RNA-dependent DNA polymerase, which will result in the formation of double-stranded cDNA from RNA. Once cDNA is made then PCR amplification follows.

We have used this kit in conjunction with other Qiagen products, in particular their RNA extraction kit (the RNeasy Kit). Our studies involve investigating and quantifying mRNA expression of peptide transporters present in porcine mucosal tissues. The cDNA produced from our samples using this kit has led to high quality material for real-time PCR analysis. Our subsequent real-time PCR data is promising and the quality of our results can be attributed to the superiority of the products we are using. In addition, the kit requires minimal additional materials, and the few materials required, such as a water bath, vortexer and microcentrifuge are normally found in most laboratories.

I recommend this kit because it is fairly easy to use. However, there are two main steps to the procedure; eliminating genomic DNA and reverse transcription, and this kit could be more convenient to the user if these steps could somehow be combined into one reaction to eliminate extra handling of samples. As well, there are always some products that are completely used up in the kit while others are not, and as a result there is some waste. In conclusion, this produce is effective although a bit pricey, but yields great results and Qiagen provides good technical support both online and in the product manual that accompanies the QuantiTect RT kit.