Lymphoma and plasma cell neoplasms
Molecular analysis
Southern blot

Author: Nikhil Sangle, M.D. (see Authors page)

Revised: 3 April 2017, last major update February 2011

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PubMed Search: southern blot lymphoma

Cite this page: Southern blot. PathologyOutlines.com website. http://www.pathologyoutlines.com/topic/lymphomamolecularsouthernblot.html. Accessed November 24th, 2017.
Definition / general
  • 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
Methods
  • 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 / cytogenetics images

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T cell receptor gene