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Analysis of Rorc Gene Rearrangement by Southern Blot Technology (CAT#: STEM-MHT-0069-LGZ)

Introduction

Official Full Name: RAR-related orphan receptor gamma<br />Also known as: TOR; Thor; Nr1f3; RORgamma<br />Enables DNA-binding transcription factor activity, RNA polymerase II-specific and transcription coactivator binding activity. Involved in several processes, including hematopoietic or lymphoid organ development; negative regulation of thymocyte apoptotic process; and regulation of gene expression. Acts upstream of or within hematopoietic or lymphoid organ development; protein phosphorylation; and regulation of gamma-delta T cell differentiation. Located in external side of plasma membrane and nucleus. Is expressed in several structures, including central nervous system; genitourinary system; hemolymphoid system; sensory organ; and thyroid gland. Used to study Sjogren's syndrome. Human ortholog(s) of this gene implicated in immunodeficiency 42. Orthologous to human RORC (RAR related orphan receptor C).




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