Poor PCR or qPCR efficiency? It can happen to everyone, even to an expert. One thing we all learn over the years about the PCR – you shall be always awake and ready for optimization and changes.
What do you usually suspect as a PCR problem number one? All PCR trouble shootings speak the same language: low PCR yield or no PCR product can be a result of different suboptimal PCR reaction parameters.
Starting from the core, one could suspect the template. DNA or cDNA can be impure, can be damaged, and can be used in too high amount or at too low concentration. Same about the primers; they might be designed wrong, synthesized and purified inappropriately or may contain contaminants that inhibit the PCR. PCR reaction conditions such as the concentration of the buffer components, dNTPs, dropped PCR polymerase activity or wrong cycling parameters may also result in too low amplification yield.
There are numerous articles and even books written about the PCR problematic. But the rule of the thumb is to set up the right control reactions and to changereaction parameters one by one. It may happen that you have to change the Taq Polymerase or dNTPs, or qPCR master mix.
flash4science is not willing to compete with all professional PCR suppliers and their trouble shootings. Instead of that, we offer sampling campaigns to try new PCR reagents and saving offers to order molecular biology tools from different life science suppliers for the best price. Scientists in UK, Germany and the US profit every day from life science saving and sampling offers with all advantages of consolidated ordering:
- flash sales – weekly offers of highly discounted molecular biology reagents or services
- flash coupons – free discount coupons to use when ordering from life science vendors
- flash samples – free exclusive samples or demos of innovative products
- long-term deals – multi-supplier life science web shop with best pricing
Communication within a network of experts is especially important in a scientific world. PCRdrive is the unique online platform for all PCR workflows. Scientists use it to share PCR-related experience, to exchange PCR protocols, to optimize PCR workflows and to solve problems together. Join PCRdrive to digitalize and accelerate your PCR.