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Analysis of Rag2 Gene Rearrangement by Southern Blot Technology (CAT#: STEM-MHT-0058-LGZ)

Introduction

Official Full Name: recombination activating gene 2
Also known as: Rag-2
Achieve multiple functions, including phosphatidylinositol phosphate binding activity; phosphatidylinositol-3,4-bisphosphate binding activity; and zinc ion binding activity. Involved in V(D)J recombination and exclusion of pre-B-cell alleles. Acts upstream or within several processes, including B cell homeostatic proliferation; lymphocyte differentiation; and positive regulation of organ growth. predicted to be in the nucleoplasm. Expected to be part of the DNA recombinase complex. Expressed in gut; hemolymphoid system; and liver. For the study of Omenn syndrome and severe combined immunodeficiency, autosomal recessive inheritance, T cell negative, B cell negative, NK cell positive. Human ortholog(s) of this gene implied in Omenn syndrome; combined cellular and humoral immune defects with granulomas; severe combined immunodeficiency; cell-positive. Orthologous to human RAG2 (recombination activating 2).




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

Gene Rearrangement Detection

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
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