Real-time PCR is a technique that monitors DNA amplification in real-time rather than at the end of the PCR run, as is the case for End-Point PCR. With the added step of fluorescent labeling, it is possible to quantify the abundance of the amplified DNA using so-called quantitative real-time PCR (qPCR). When qPCR is used in combination with reverse transcription and cDNA synthesis, this is called RT-qPCR, a common technique for the analysis of gene expression.
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How does Real-Time PCR work?
Here at ZAGENO, you can find kits for different templates, such as:
Real-time PCR (qPCR) kits will provide polymerases, oligonucleotides, and buffers for your experiments. Before comparing kits from different suppliers on our product page, though, there are a few factors to consider:
You should keep in mind the size of your amplicon. While in general qPCR amplifies short fragments of around 150 bases, there are polymerases with higher processivity. A side note: if you plan on performing Hot Start qPCR (prevents non-specific amplification at room temperature), look for kits including enzymes suitable for this application.
If you expect to amplify a template present in low quantities in your sample, consider kits that use SYTO 9 as an intercalating dye, which has higher sensitivity. For standard applications, SYBR Green I is still an excellent choice.
Alternatively, probe-based qPCR, use labeled fluorescent probes instead of intercalates. In doing so provides excellent sensitivity for sequence–specific detection, but need to be prepared specifically for your template beforehand.
If you are using RT-qPCR, make sure to pick a one- or two-step kit, according to your experimental setup.
ZAGENO's Real-Time PCR Troubleshoot will help guide you through any diffculties encountered.
Compare Real-Time PCR Kits!
With our compare function, you can avoid all the time and energy wasted sifting through multiple web pages from different suppliers. At ZAGENO you can clearly see kits side-by-side, with the relevant attributes for each kit neatly in line for easy selection of the best product for you.