PCR primers and conditions used for reverse transcriptase–PCR analysis
Gene . | Primers . | PCR condition, 35 cycles . |
|---|---|---|
| Rag-2 | 5′-CAATACCTGGGTTCCGTTCTATT-3′; 5′-GCTCTAGAATTAATCAAACAGTCTTCTAAGG-3′ | 94°C, 30 s; 45°C, 30 s; 72°C, 30 s |
| κ germline transcript | 5′-CCACATGCCTTTCTTCAGGGACAAGTGGGA-3′ (κ GT-5′); 5′-GTTATGTCGTTCATACTCGTCCTTGGTCAAC-3′ (κ GT-3′) | 94°C, 1 min; 64°C, 30 s; 72°C, 30 s |
| Spi-B | 5′-GTCCTAACCCCTCCACCTATAG-3′; 5′-TAGGAGCAACCCCAGCAAGAAC-3′ | 94°C, 1 min; 64°C, 30 s; 72°C, 30 s |
| Irf-4 | 5′-ACAGGATTGTTCCAGAGGGAGC-3′; 5′-CGAGGATGTCCCGGTAATACAG-3′ | 94°C, 1 min; 64°C, 30 s; 72°C, 30 s |
| λ5 | 5′-ATGAAGCTCAGAGTAGGACAGAC-3′; 5′-CTAAGAACACTCAGCAGGTGA-3′ | 94°C, 1 min; 61°C, 30 s; 72°C, 1 min |
| Vpre-B | 5′-CAGGTCTAGAGCCTGGCCTGGACGTCTG-3′; 5′-GTCTGAATTCCTCCAGACCCTAAGATCCC-3′ | 95°C, 1 min; 55°C, 30 s; 72°C, 30 s |
| κ mature mRNA | 5′-GGCTGCAGSTTCAGTGGCAGTGGRTCWGGR-3′ (FWR3); 5′-ACGCCATTTTGTCGTTCACTGCCA-3′ (Cκ-3′) | 95°C, 1 min; 61°C, 30 s; 72°C, 1 min |
| E2A | 5′-AGGCACTGGCCTCGATCTAC-3′; 5′-TGGCCAGGCAAAAGCCCATG-3′ | 95°C, 1 min; 60°C, 30 s; 72°C, 30 s |
| PKCη | 5′-CCATGACTGAGAAGAGGATC-3′; 5′-AATGTAGTCAGGTGTACCGC-3′ | 95°C, 1 min; 56°C, 1 min; 72°C, 1 min |
| HPRT | 5′-TTGCTGGTGAAAAGGACCTCTCG-3′; 5′-CCACAGGACTAGAACACCTGCTAA-3′ | 95°C, 1 min; 55°C, 1 min; 72°C, 1 min |
Gene . | Primers . | PCR condition, 35 cycles . |
|---|---|---|
| Rag-2 | 5′-CAATACCTGGGTTCCGTTCTATT-3′; 5′-GCTCTAGAATTAATCAAACAGTCTTCTAAGG-3′ | 94°C, 30 s; 45°C, 30 s; 72°C, 30 s |
| κ germline transcript | 5′-CCACATGCCTTTCTTCAGGGACAAGTGGGA-3′ (κ GT-5′); 5′-GTTATGTCGTTCATACTCGTCCTTGGTCAAC-3′ (κ GT-3′) | 94°C, 1 min; 64°C, 30 s; 72°C, 30 s |
| Spi-B | 5′-GTCCTAACCCCTCCACCTATAG-3′; 5′-TAGGAGCAACCCCAGCAAGAAC-3′ | 94°C, 1 min; 64°C, 30 s; 72°C, 30 s |
| Irf-4 | 5′-ACAGGATTGTTCCAGAGGGAGC-3′; 5′-CGAGGATGTCCCGGTAATACAG-3′ | 94°C, 1 min; 64°C, 30 s; 72°C, 30 s |
| λ5 | 5′-ATGAAGCTCAGAGTAGGACAGAC-3′; 5′-CTAAGAACACTCAGCAGGTGA-3′ | 94°C, 1 min; 61°C, 30 s; 72°C, 1 min |
| Vpre-B | 5′-CAGGTCTAGAGCCTGGCCTGGACGTCTG-3′; 5′-GTCTGAATTCCTCCAGACCCTAAGATCCC-3′ | 95°C, 1 min; 55°C, 30 s; 72°C, 30 s |
| κ mature mRNA | 5′-GGCTGCAGSTTCAGTGGCAGTGGRTCWGGR-3′ (FWR3); 5′-ACGCCATTTTGTCGTTCACTGCCA-3′ (Cκ-3′) | 95°C, 1 min; 61°C, 30 s; 72°C, 1 min |
| E2A | 5′-AGGCACTGGCCTCGATCTAC-3′; 5′-TGGCCAGGCAAAAGCCCATG-3′ | 95°C, 1 min; 60°C, 30 s; 72°C, 30 s |
| PKCη | 5′-CCATGACTGAGAAGAGGATC-3′; 5′-AATGTAGTCAGGTGTACCGC-3′ | 95°C, 1 min; 56°C, 1 min; 72°C, 1 min |
| HPRT | 5′-TTGCTGGTGAAAAGGACCTCTCG-3′; 5′-CCACAGGACTAGAACACCTGCTAA-3′ | 95°C, 1 min; 55°C, 1 min; 72°C, 1 min |
Names of the primers depicted in Figure 2C are in parentheses