Comparison Of Effects Of Different Buffers For Dna Fragment Elution From Agarose Gel

Abstract

Purification of nucleic acids is an essential procedure for most experiments in molecular biology because the presence of impurities can interfere with such molecular applications as restriction digestion, ligation, cloning, labeling sequencing, PCR and RT-qPCR. In order to solve this problem, several methods for purification of DNA fragments have been developed, such as those utilizing phenol-chloroform, paper strip and magnetic particles (for aqueous solutions); other methods include purifying DNA from fragments of agarose gel, known as the freeze-squeeze method. This method has been widely used for DNA purification but has the disadvantage of using low-melting point agarose or requiring high concentrations of DNA samples. Based on the above mentioned considerations and our needs for purifying and analyzing small fragments of DNA or molecules. Moreover, there are many kinds of buffer which are used as commercial kits buffers such as KI, Guanidine Hydrochloride (GuHCl), Guanidine Thyocynate (GuSCN), etc. In fact, it is important to emphasize that this thesis is to compare these buffer effects to the task. After go through many steps we gained the result that KI and GuSCN had as similar quality as commercial buffer. Although, KI always had the outstanding result when measured by NANODROP the out-come expressed by gel electrophoresis is more reliable than by NANODROP measurement. Therefore, GuSCN is the best one.