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| Status |
Public on Jul 16, 2013 |
| Title |
High throughput sequencing of immunoglobulin (Ig) heavy (IgH) and light (IgL) chain repertoires |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
We used 454 sequencing to assess the repertoire of B cell subsets from bone marrow, spleen, and small intestinal lamina propria from two mouse strains. We used a RAG2-GFP reporter mouse strain (129Sve background) to isolate CD19+ RAG2+ B lineage cells from bone marrow and small intestinal lamina propria and total splenic B cells. We used 5' RACE to amplify cDNA libraries using primers specific for the mu constant region of IgH and the Ig kappa constant region. We also used this technique to analyze total B cell libraries from Swiss Webster germ-free mice to compare to littermate controls that were cohoused with regular specific pathogen free (SPF) mice for 7 days.
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| Overall design |
Examination of the Ig kappa repertoire and IgH repertoire in RAG2+ bone marrow B lineage cells compared to RAG2+ small intestinal lamina propria B lineage cells or total splenic B cells. There are 8 (Ig kappa) or 4 (IgH) independent experiments comparing repertoires in RAG2-GFP mice. Each experiment in RAG2-GFP+ mice consisted of a pool of 8-12 mice. There are 3 experiments comparing germ-free to colonized mouse total B cell repertoires, each consisting of one mouse per condition.
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| Contributor(s) |
Wesemann DR, Portuguese AP, Meyers RM, Gallagher MP, Cluff-Jones K, M.Magee J, Panchakshari RA, Rodig SJ, Kepler TB, Alt FW |
| Citation(s) |
23965619 |
| Submission date |
Jul 15, 2013 |
| Last update date |
May 15, 2019 |
| Contact name |
Frederick Alt |
| Organization name |
Boston Children's Hospital
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| Department |
Program in Cellular and Molecular Medicine
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| Street address |
1 Blackfan Circle
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| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02155 |
| Country |
USA |
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| Platforms (1) |
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| Samples (40)
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| Relations |
| BioProject |
PRJNA212030 |
| SRA |
SRP027381 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE48870_IgH_Col_LP.txt.gz |
889 b |
(ftp)(http) |
TXT |
| GSE48870_IgH_GF_LP.txt.gz |
1008 b |
(ftp)(http) |
TXT |
| GSE48870_IgH_RAG2_BM.txt.gz |
1.0 Kb |
(ftp)(http) |
TXT |
| GSE48870_IgH_RAG2_LP.txt.gz |
1.1 Kb |
(ftp)(http) |
TXT |
| GSE48870_IgH_total_spl.txt.gz |
1.0 Kb |
(ftp)(http) |
TXT |
| GSE48870_Igk_Col_LP.txt.gz |
602 b |
(ftp)(http) |
TXT |
| GSE48870_Igk_GF_LP.txt.gz |
629 b |
(ftp)(http) |
TXT |
| GSE48870_Igk_RAG2_BM.txt.gz |
1.0 Kb |
(ftp)(http) |
TXT |
| GSE48870_Igk_RAG2_LP.txt.gz |
1008 b |
(ftp)(http) |
TXT |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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