How to Optimize Emulsification PCR

In less than thirty years, the polymerase chain reaction(PCR) has swept the life science research, becoming the essential experimental technology of many biological laboratories. Although the basic premise is the same—amplification purposes DNA samples, in recent years, there have been a lot of innovations to put the technology into a number of application areas.

In order to achieve these objectives, the researchers used a high throughput PCR technology in an emulsion solution. These droplets, often taking over only a few liters of volume, can maximize the polymerase reagent ratio, and reduce the waste caused by large volume PCR reaction, which greatly improves the efficiency of work.

Currently, a lot of digital PCR instruments can do such experiments on the market. One of the merits is that such a large number of independent reactions minimize the cross contamination by product. More importantly, the emulsion PCR method can make the tedious experiment evolve into a high-throughput expanded experiment. However, this approach is also flawed. Scientists are still looking for a better solution.

Optimizing aptamer

Selex aptamer is screened out from a random oligonucleotide library through in vitro SELEX technology. Oligonucleotide sequences have high binding specificity with the target molecules, so designing aptamer is often a thankless job. It's quite time-consuming to find suitable aptamer from a large number of possible sequences, which requires to clone the aptamer sequence into the plasmid, then cultures bacteria and selects appropriate analysis cloning. All those candidate aptamer is needed for sequencing, and it is is not cheap. Then it is needed to compare the sequences to find the good aptamer. Finally, the researchers also need to analyze the binding of each aptamer separately. Those will undoubtedly spend a lot of manpower and financial resources.

In order to solve this problem, a PCR based on agarose is used. The agarose droplet itself can work like beads by adjusting the temperature to manipulate agarose from wet/soft nature solid. Therefore, the agarose droplet can facilitate the PCR reaction, and also can be arrayed as solid micro beads.