Nucleic acids detection by dot-blot method

Dot-blot is one of biological methods that are normally used in research laboratories and especially in the diagnostics. It is the most commonly used method for identification and immunodetection of particular proteins which may be markers of various diseases. The main aim of the experiment was to develop a simple, inexpensive and rapid method for specific detection of nucleic acids, especially DNA, and then this procedure apply to the detection of DNA modified by platinum cytostatic drugs. Despite platinum cytostatic drugs‘ common use in chemotherapy of various cancer types, their biochemical effect is still not completely clear. The generally accepted opinion is that the drug binds to cellular DNA. It was observed that cisplatin is bound to the DNA the most compared to oxaliplatin and carboplatin.

Fig. 1 Concentration series of DNA

Tab. 1 Convert concentration on weight DNA in 2 ml of sample

Figure 1 Dependence of color intensity spot on the amount of DNA (inset - linear portion of the curve)

Fig. 2 The intensity of blotting membranes depending on the length blocks and using blocking solution

Fig. 3 Dot-blot platinum DNA adducts

Figure 2 Dependence of color intensity of the spot on the quantity and type of the applied cytostatic drug

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