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Analysis of Echinococcus Granulosus of Spanish Origin by Southern Blot Technology (CAT#: STEM-MHT-0123-LGZ)

Introduction

Cloned DNA segments have been used as DNA probes in restriction endonuclease analysis and Southern blot hybridization to characterize E. granulosus material from Spain. Two genetic variants-a sheep/cattle/human form, very similar to the United Kingdom sheep strain, and a parasite from donkeys, genetically similar to the United Kingdom horse strain—have been identified using this approach.




Principle

Under certain conditions, two single strands of nucleic acid with certain homology can be specifically hybridized to form double strands according to the principle of base complementarity. Generally, DNA molecules to be detected are digested with restriction enzymes, separated by agar-gel electrophoresis, denatured and transferred to nitrocellulocellulose film or nylon film or other solid phase support according to their position in the gel, fixed and then reacted with DNA probes labeled with isotopes or other markers. This is followed by autoradiography or an enzyme reaction to detect the amount of specific DNA molecules. If the object to be tested contains a sequence that is complementary to the probe, the two are combined by the principle of base complementarity, and the free probe is washed and detected by self-development or other suitable techniques, thus revealing the fragment to be tested and its relative size.

Applications

DNA Analysis

Procedure

1. Sample Processing
2. DNA Extraction and Digestion
3. Gel Electrophoresis
4. Gel Pretreatment
5. Transfer membrane
6. Probe Labeling
7. Prehybridization (blocking)
8. Southern hybridization
9. Membrane washing
10. Autoradiographic Assay
11. Results Analysis

Materials

Sample: DNA, Bacterial Fluid/Tissue/Cell