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Lymphoma - B cell neoplasms

Molecular analysis - Southern blot


Reviewer: Nikhil Sangle, M.D., University of Utah and ARUP Laboratories (see Reviewers page)
Revised: 26 February 2011, last major update February 2011
Copyright: (c) 2001-2011, PathologyOutlines.com, Inc.

General
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● Detects DNA via restriction endonucleases, electrophoresis and probes
● For lymphoma, detects alterations of DNA restriction fragment length from rearrangement of immunoglobulin and T cell receptor genes
● Rearrangement forms a distinct band if only 1% of total cells are affected, so it is very sensitive; only a single germline band is present if polyclonal or not clonal

Methodology
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● Difficult and slow to perform
● Usually performed to analyze IgH and TCR-beta for clonality
● Procedure:
● (1) Extract DNA
● (2) Cut DNA into small fragments with restriction enzymes (EcoRI, HindIII, BamHI)
● (3) Apply DNA to electrophoretic gel (agarose)
● (4) Apply nylon membrane (blot) over gel to transfer DNA fragments onto membrane
● (5) Denature the probe DNA and DNA fixed to the blot to allow hybridization; then add probe DNA
● (6) Detect the probe (by xray if probe is radioactive)

Molecular images
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T cell receptor gene

End of Lymphoma - B cell neoplasms > Molecular analysis - Southern blot


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