Alexander Y. Liu, Hiroto Koga, and Chihiro Aiya, members of the Kyoto University undergraduate research team iGEM Kyoto, have developed a new reaction mixture for DNA cloning. Just by mixing two enzymes, the cost is one thousandth of expensive reagents.Performance was similar to that of commercial products.
DNA cloning is a core technology of molecular biology, and requires a complicated process using "scissors" and "glue" to cut DNA.Recently, a simple method based on "homologous recombination" has been used, but the need for expensive reagents puts pressure on the budgets of many laboratories.
The research team was hesitant to buy expensive reagents due to a small budget, and started research by thinking, ``Can we create a new reaction solution ourselves?''Focusing on reports that "homologous recombination" can be achieved by using "SLiCE", a solution extracted from Escherichia coli using a special method, without using expensive reagents. challenged.
Studies have shown that two enzymes, exonuclease III (ExoIII) and exonuclease T (ExoT), are responsible for the activity. By simply mixing two enzymes, we succeeded in completely reproducing the activity of SLiCE, in which countless proteins are dissolved.ExoIII, which is responsible for the main activity, was purchased commercially, but since it is diluted and used, if you buy a reagent of 2 yen, you can perform 2 recombination reactions (commercially available kits cost about 80 yen per reaction). ).It has become possible to make cocktails (mixtures) that are comparable to expensive kits at a very low cost.
The prepared cocktail could be stored under very stable conditions, and the activity did not decrease even after long-term storage.This achievement is expected to lead to a significant reduction in the cost of DNA cloning around the world.
Paper information:[Genes to Cells] Quick and affordable DNA cloning by reconstitution of Seamless Ligation Cloning Extract using defined factors
