Table 2. PCR and sequencing primers for the analysis of the human CLCNKB gene.
| Exon/ Promoter | Forward primer (5´- 3´) | Reverse primer (5´- 3´) | Annealing T (°C) | Amplicon Length (bp) |
|---|---|---|---|---|
| Promoter | * CTCTCCCCATTCACAGGGTG | * TGGACAGGTGTGTGTTCCAA | 56,3 | 966 |
| † CGGCCTCCGTGATCTTAGAC | † AGCATGACCACAGCCTCC | |||
| 1 | * CTCTGTGCAGCTATGGTGGG | * CTGTCCACCTATGAGCACCC | 56.3 | 429 |
| 2 | ‡ ACTGGAAGGGCCTAGAGGCAGT | ‡ GATGTCCTGAGTGGTCCTCCAG | 60 | 231 |
| 3 | § CACTGTGTCACCACTGTCACC | § AGGAGTAAAGCCAGGACCAGA | 60 | 460 |
| FD | || ¶ TGCCCCACCCTGTGCCGTGAC | || ¶ GGGTGGTTGGGATGCCCTCAC | 66,8 | 2558 |
| 4 | || # GAGGCTGTGGGTGCCTCCCTG | || # AGTGGGGACTGGCGTAGCGAC | 61,4 | 200 |
| 5-7 | § AGATCTTGTCCCCAAAGGAAA | § GGCTGAAGTGAGAACTAGAATGA | 60 | 950 |
| † AGTGGTATGGGCAGGGGT | † AGGTAGGCAGCCATCATCAC | |||
| † CACCTGTCTGTGATGATGG | † GGGTAGGGTGGTTGGGA | |||
| FE | || ¶ CCCTCCTGGCCCTGCCCAC | || ¶AGCTCGCTGAGAGGTCCCCAG | 66,8 | 2082 |
| 8 | || # GGAGGGCCCACCTGAGATCAG | || # GCAGGGCCAGGGTCAGGCAG | 61,4 | 193 |
| 9-10 | || # CGCCATCTTGGCTCCCCACTG | || # AGCTCGCTGAGAGGTCCCCAG | 61,4 | 374 |
| 11-12 | § CTGACCCCACAGGTTCTGT | § ** CCAGGGCAGAGGTTAGAGGC | 60 | 1099 |
| † GCCTCCTTTGCGTGTAT | † TAACCAGGAAGAAGGCAAGG | |||
| † GGTTCACCATCTTTGGGA | † CTGACCTCCCGAAGCTGTAG | |||
| FF | || ¶ GTCGGGCTCTGGGCTCATGTC | || ¶ CAGTCAGCCTGAGGTGGGCAC | 66,8 | 2660 |
| 13-14 | || # TCTAGGACACTCCCCTGTCCC | || # CCCTGGGGAACCACCAGCCAA | 61,4 | 519 |
| 15 | †† CATCACTCCCTCGTGGCTCCTG | †† CTACGGTGGCGTTTCTTTTTCG | 60 | 518 |
| 16 | †† GCTAAAGTGGAGCTGGTCTG | †† GCAACAAGGATTTGGAGG | 60 | 750 |
| † CCACAGCATCACCACACT | † CACAACCTTGACCACCTCCT | |||
| 17-18 | § CACAATAGCCCCATAGGAACA | § CTCTCCCACTTCCCTCATCTC | 60 | 672 |
| † GGTCAGAGAGAGGCATCCTG | † ATGTCCTGGAGACACTGCTG | |||
| 19 | || GGGCACCTTCTACCCTCCAGTG | || GTCTTCTCAGGCATAGGTTCCCTG | 60 | 187 |
| 20 | || CTACATCCCCCCGCACCACCAC | || AGGGTCTCAGCCCAACCTC | 60 | 153 |
According to DNA sequence (Ensembl: ENST00000375679); Bases that differs from the CLCNKA gene (Ensembl: ENST00000331433) are marked in bold; * Newly designed primers; † Newly designed internal primers for sequencing; ‡ Previously designed primers [20]; § Previously designed primers [21]; || Primers kindly provided by the Hôpital Européen Georges Pompidou, Service de Génétique, Paris; ¶ Specific primers for long-range PCR; # Internal specific primers for nested PCR; ** Reverse primer modified (last base removed because it is a SNP); †† Previously designed primers [22].