ABSTRACT

It has been shown that chemical and/or mechanic pre-treatments increase the success rate of DNA isolation before applying the kit procedures in commercial kits. It has been observed that RMY preprocess contributes positively to DNA isolation from Blastocystis sp. isolates

As a result of agarose gel visualization out of 12 samples, we have observed the 1100 bp band belong to Blastocystis sp. in 11 samples (91.6%) with RMY and in 6 samples (50%) with SGM. It was detected that the bands were more visible and prominent with RMY. Meanwhile, it has been determined that RMY technique is practical and it can be performed in a short time

Blastocystis sp. was determined with native-lugol method in the12 stool samples that were sent to laboratory for routine parasitological examination and the duplicate cultures were maintained in Ringer's solution containing 10% horse serum and 0.05% asparagine. Before DNA isolation, culture suspensions were processed in Ringer’ solution the technique that we have modified (RMY) and sucrose gradient method (SGM). Upon two different pre-treatments, DNA isolation was performed with DNAzol® (MRC,USA) according to manufacturer’s protocol and instructions. Then, 1100 bp gene belong to protozoa SSUrDNA locus was amplified with F1 and RB primers by PCR technique. Amplicons were runned and observed on 1.5% agarose gel

In the field of research and diagnosis of protozoa, the first step of the molecular methods is to isolate the DNA of protozoa. DNA isolation from stool samples has difficulties due to the complex and particulate structure of the stool and the presence of inhibitors. In this study, before isolating the DNA from Blastocystis sp. isolates, we aimed to examine the effects of the two different preprocesses applied before the isolation