Abstract:

Acid guanidium phenol chemicals such as Trizol allow high-quality total RNA isolation from many samples. In order to be able to make high-quality RNA isolation, with an economic and repeatable method from wooden plasentom (total, maternal and fetal), the total RNA is centralized with an acid solution containing 1 ml of guanidine tiyosyanate, sodium acetate, phenol and chlorform. Total RNA is disintegrated with isopropanol and no purification kit has been used. The isolation is done according to two different protocols. The isopropanol phase is performed at (a) room temperature (20-22°C) and (b) on ice (0-4°C). The RNA quality is made spectraphotometrically, with optical dancite (OD) measurement (OD260 nm/OD280 nm > 1.7). The integrity of the RNA is controlled by agarose gel electrophoresis. According to the electrophoresis results (a) according to the protocol 28s, 18s and 5s rRNA bands were observed, while (b) according to the protocol often degrade RNA bands were observed. The total RNA concentration (b) is significantly high according to the protocol, while (a) the purity and integrity according to the protocol has been achieved very good quality RNA. PCR is made by making the cDNA conversion from the total RNA obtained. RT-PCR products are successfully obtained from the RNA isolated according to the (a) protocol. (a) the protocol indicates that cow plasentom is an economic and repeatable method for high-quality RNA isolation from total, maternal and fetal tissues, without using any purification kit;

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