A reliable approach for sequencing clone-specific CDRIII regions in B-cell lymphoma.
Authors
Straka, CPettengell, Ruth
Pielmeier, A
Cross, Michael A
Emmerich, B
Crowther, Derek
Testa, Nydia G
Dexter, T Michael
Affiliation
Department of Experimental Haematology, C.R.C. Paterson Institute for Cancer Research, Manchester, UK.Issue Date
1994
Metadata
Show full item recordAbstract
We report a reliable approach for sequencing lymphoma-specific CDRIII regions. CDRIII regions present in DNA prepared from routinely fixed and paraffin-embedded diagnostic lymph node material were amplified by the use of consensus VH and JH primers via PCR. PCR products were subcloned directly, without purification or modification of PCR fragments. Only small amounts of miniprep plasmid DNA of recombinant clones were required for cycle sequencing, resulting in autoradiograms of high quality. The easy and reproducible method which we describe has enabled us to determine the lymphoma-specific CDRIII region in 7/11 high-grade non-Hodgkin's lymphomas as well as in 3/3 cases of ALL and 1/1 case of a centroblastic/centrocytic lymphoma. The obtained sequence data can serve to generate lymphoma-specific oligonucleotides, which then can be used as PCR primers or hybridization probes for the detection of minimal residual disease in individual patients.Citation
A reliable approach for sequencing clone-specific CDRIII regions in B-cell lymphoma. 1994, 5 Suppl 1:79-84 Ann. Oncol.Journal
Annals of OncologyDOI
10.1093/annonc/5.suppl_1.S79PubMed ID
8172824Type
ArticleLanguage
enISSN
0923-7534ae974a485f413a2113503eed53cd6c53
10.1093/annonc/5.suppl_1.S79
Scopus Count
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