A reliable approach for sequencing clone-specific CDRIII regions in B-cell lymphoma.

2.50
Hdl Handle:
http://hdl.handle.net/10541/97346
Title:
A reliable approach for sequencing clone-specific CDRIII regions in B-cell lymphoma.
Authors:
Straka, C; Pettengell, Ruth; Pielmeier, A; Cross, Michael A; Emmerich, B; Crowther, Derek; Testa, Nydia G; Dexter, T Michael
Abstract:
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.
Affiliation:
Department of Experimental Haematology, C.R.C. Paterson Institute for Cancer Research, Manchester, UK.
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 Oncology
Issue Date:
1994
URI:
http://hdl.handle.net/10541/97346
DOI:
10.1093/annonc/5.suppl_1.S79
PubMed ID:
8172824
Type:
Article
Language:
en
ISSN:
0923-7534
Appears in Collections:
All Christie Publications ; All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorStraka, Cen
dc.contributor.authorPettengell, Ruthen
dc.contributor.authorPielmeier, Aen
dc.contributor.authorCross, Michael Aen
dc.contributor.authorEmmerich, Ben
dc.contributor.authorCrowther, Dereken
dc.contributor.authorTesta, Nydia Gen
dc.contributor.authorDexter, T Michaelen
dc.date.accessioned2010-04-26T10:04:46Z-
dc.date.available2010-04-26T10:04:46Z-
dc.date.issued1994-
dc.identifier.citationA reliable approach for sequencing clone-specific CDRIII regions in B-cell lymphoma. 1994, 5 Suppl 1:79-84 Ann. Oncol.en
dc.identifier.issn0923-7534-
dc.identifier.pmid8172824-
dc.identifier.doi10.1093/annonc/5.suppl_1.S79-
dc.identifier.urihttp://hdl.handle.net/10541/97346-
dc.description.abstractWe 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.en
dc.language.isoenen
dc.subject.meshBase Sequence-
dc.subject.meshCloning, Molecular-
dc.subject.meshHumans-
dc.subject.meshImmunoglobulin Variable Region-
dc.subject.meshLymphoma, B-Cell-
dc.subject.meshMolecular Sequence Data-
dc.subject.meshReproducibility of Results-
dc.titleA reliable approach for sequencing clone-specific CDRIII regions in B-cell lymphoma.en
dc.typeArticleen
dc.contributor.departmentDepartment of Experimental Haematology, C.R.C. Paterson Institute for Cancer Research, Manchester, UK.en
dc.identifier.journalAnnals of Oncologyen

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