Dna Slot Blot Protocol

  
  1. Dna Slot Blot Protocol Definition

Slot blot quantitation can help determine whether the DNA is human or non-human. DNA quantitation must be accomplished on all samples by yield gel and/or slot blot before amplification. Slot blot is required for all unknown samples. It is not required for standard samples. The decision to use slot blot on standard samples is left to the. Four different sample number, size and style of Hybridisation Manifold are offered; two types of Dot Blotter and two types of Slot Blotter. Typical applications include clone screening with DNA/RNA probes in Southern/Northern blots and immunological screening with antibodies in Western blots.

A Southern blot is a way to analyze DNA molecules. The protocol was developed by Edward Southern. And if you were going to perform a Southern blot, you would first want to separate DNA based upon size in a gel along an electric field.

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.

Dna Slot Blot Protocol
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 Definition

1. Scitable by nature education; Northern blot; [online]. [cit. 2014-01-20] Dostupné z: http://www.nature.com/scitable/definition/northern-blot-287.
2. Scitable by nature education; Southern blot; [online]. [cit. 2014-01-20] Dostupné z: http://www.nature.com/scitable/definition/southern-blot-289.
3. Dot Blot Analysis; Teacher's Guidebook; [online]. [cit. 2014-01-20] Dostupné z: http://www.gbiosciences.com/PDF/Protocol/502_Dot_Blot_Analysis_TEACHER.pdf.
4. Ceppellini R., Polli E., Celada F.: Proceedings of the Society for Experimental Biology and Medicine, 96, 572 (1957).
5. Rothfiel.Nf, Stollar B. D.: Journal of Clinical Investigation, 46, 1785 (1967).
6. Schur P. H., Sandson J.: New England Journal of Medicine, 278, 533 (1968).
7. Tojo T., Friou G. J.: Science, 161, 904 (1968).
8. Fuertes M. A., Castilla J., Alonso C., Perez J. M.: Curr. Med. Chem., 10, 257 (2003).
9. Wang D., Lippard S. J.: Nat. Rev. Drug Discov., 4, 307 (2005).
10. Krizkova S., Adam V., Eckschlager T., Kizek R.: Electrophoresis, 30, 3726 (2009).