The planning and Construction of different DNA markers using Plasmids and Lambda DNA

DNA markers are one of the most important indicators for estimating Molecular weight of DNA samples, although it used in widespread medical and research laboratories. These markers are very divers and have been prepared in different manners and from different sources of DNA. But unfortunately, DNA markers havent been made in our country and all of the markers that we use are made in a foreign country.The aim of this research is settings a suitable technology to produce this product in the lab. With this aim, we used two different strains of lambda: c1857sam7 and EMBL3A both of which are lytic phages as a DNA source. These were grown in the suitable host, after plaque appearance on the bacterial lawn, suitable titer for phage collecting was determined. We also optimized plasmid purification method for extraction of pBR322, pUC18 and recombinant VZV plasmid DNA and designed fragments in the markers have been constructed by digesting these DNAs with variant enzymes. In this study, we made seven DNA markers out of which four of them were made for the first time in the world (λ/Hind III/BamH1, λ/Hind III/EcoR1, Sam2, Sam1) and although foreign models of three of them exist but they were made in our country for the first time (λ/Pst I, λ/Hind III, pBR332/MspI). The other goal of this study was to determining the best conditions for maintaining and preserving these markers in the lab which was successfully performed.