Molecular cloning of IGλ rearrangements using long-distance inverse PCR (LDI-PCR)
Article first published online: 4 DEC 2012
© 2012 John Wiley & Sons A/S
European Journal of Haematology
Volume 90, Issue 1, pages 59–67, January 2013
How to Cite
Shimanuki, M., Sonoki, T., Hosoi, H., Watanuki, J., Murata, S., Kawakami, K., Matsuoka, H., Hanaoka, N. and Nakakuma, H. (2013), Molecular cloning of IGλ rearrangements using long-distance inverse PCR (LDI-PCR). European Journal of Haematology, 90: 59–67. doi: 10.1111/ejh.12037
- Issue published online: 14 DEC 2012
- Article first published online: 4 DEC 2012
- Accepted manuscript online: 1 NOV 2012 03:32AM EST
- Manuscript Accepted: 26 OCT 2012
- Ministry of Education, Culture, Sports, Science, and Technology (MEXT)
Figure S1. (A) Enzyme map of IGλ locus, isotype-specific probes and positions of primer sets. This figure is based on a previous report (14) and published sequence data (accession no. X51755 and NG_000002). B: BamHI, E: EcoRI, G: BglII, H: HindIII, S: SphI, and T: TaqI. The IGLJ2 probe recognizes both λ 2 and λ 3 segments (indicated by an asterisk). IGLC2D contains an SphI site. Arrowheads indicate primer pairs. (B) Schema of deregulation of the partner gene by IGλ translocation is illustrated. The partner gene is affected by IGλ from telomeric sides. (C) Germline fragments of eachλ segment can be amplified by LDI-PCR using respective primer sets. 1: λ1 (11 kb of Bgl II fragment amplified with lambda 1 primer set), 2: λ2 (8.1 kb of Hind III fragment amplified with lambda 2a primer set), 3: λ2 (7.8 kb of Hind III fragment amplified with lambda 2b primer set), 4: λ3 (3.7 kb of Taq I fragment amplified with lambda 3a primer set), 5: λ3 (4.1 kb of Taq I fragment amplified with lambda 3b primer set), 6: λ4 (3.5 kb of BamH I fragment amplified with lambda 4 primer set), 7: λ5 (4.4 kb of Bgl II fragment amplified with lambda 5 primer set), 8: λ6 (4.4 kb of BamH I fragment amplified with lambda 6 primer set), 9: λ7 (1.9 kb of BamH I fragment with lambda 7 primer set). D: DNA marker (λ/HindIII and λ/HaeIII digest).
Table S1. Primers for LDI-PCR.
Table S2. DNA size of germ line fragment.
Table S3. Primers for verification.
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