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Analysis of LARGE1 Gene Rearrangement by Southern Blot Technology (CAT#: STEM-MHT-0103-LGZ)

Introduction

Official Full Name: LARGE xylosyl- and glucuronyltransferase 1
Also known as: LARGE; MDC1D; MDDGA6; MDDGB6
This gene encodes a member of the n-acetylglucosamine transferase gene family. It encodes a glycosyltransferase involved in the glycosylation of α-triglycan glycans and the synthesis of glycoproteins and sphingolipid sugar chains. It may also be involved in the addition of repeating disaccharide units. The protein encoded by this gene is a glycotransferase that adds final xylose and glucuronic acid to betatriuronic acid, thereby allowing betatriuronic acid to bind ligands, including laminin 211 and neurotoxin . Mutations in this gene cause multiple forms of congenital muscular dystrophy characterized by cognitive impairment and abnormal glycosylation of alpha-dystrophy. Alternative splicing of this gene results in multiple transcript variants encoding the same protein.




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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