Dna Slot Blot Protocol



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The transfer of macromolecules such as nucleic acids and proteins to solid-phase membranous support is known as blotting. Fragments of DNA and RNA molecules separated by gel electrophoresis are transferred to a nylon or nitrocellulose membrane in a process termed as Southern and Northern blotting, respectively. Abstract: This protocol describes how to prepare a DNA dot blot with a dot blot apparatus and a vacuum manifold. Author: Simon Dawson. Regularly it is useful to evaluate the abundance of certain RNA or DNA in the extracted nucleic acid mixture by dot blot or slot blot hybridisation without prior digestion and electrophoresis.

Slot

Marcela Vlcnovska, Kristyna Smerkova, Marketa Vaculovicova, Sona Krizkova, Rene Kizek

Dot-blot is one of biological methods that are normally used in research laboratories and especially inthe diagnostics. It is the most commonly used method for identification and immunodetection of particularproteins which may be markers of various diseases. The main aim of the experiment was to developa simple, inexpensive and rapid method for specific detection of nucleic acids, especially DNA, and thenthis procedure apply to the detection of DNA modified by platinum cytostatic drugs. Despite platinumcytostatic drugs‘ common use in chemotherapy of various cancer types, their biochemical effect is still notcompletely clear. The generally accepted opinion is that the drug binds to cellular DNA. It was observedthat 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
Dna Slot Blot Protocol

Dna Slot Blot Protocol Definition

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