Sequence and position of primers used for RT-PCR, genotyping, and mutagenesis
| Primer*name . | Nucleotide sequence† . | Nucleotide position‡ . | PCR-product . |
|---|---|---|---|
| cDNA | |||
| SP-1 | 5′-GAGCCAGAGGAAGAGATAGCC-3′ | −21 to −1 | |
| AS-2 | 5′-GAAAAACAGCAGTTTGCTGAGACC-3′ | 1260 to 1237 | |
| AS-3 | 5′-GGAAGTTAGCTGGAGCAGTCAGGC-3′ | 1234 to 1211 | −21 to 1234 |
| genomic-DNA | |||
| SP-4 | 5′-ctgtgtctcttgccccacaa-3′ | 20 to 1 up Ex6 | |
| AS-5 | 5′-caactcccattccctccttctgcc-3′ | 72 to 49 down Ex6 | Int5-Int6 (221 nt) |
| SP-6 | 5′-GGCGTTGTCAATGTACCTTTCAGCC-3′ | 546 to 570 | |
| SP-7 | 5′-TCCTGGTCTGACCTCAGTGCCCTG-3′ | 637 to 660 | |
| AS-8 | 5′-tgcgtatcttgcagagcc-3′ | 49 to 32 down Ex7 | Ex7-Int7 (76 nt) |
| SP-9 | 5′-GACTCAGTCTTTCAGCCCC-3′ | 812 to 830 | |
| AS-10 | 5′-CCATTGCAATGCAGGCCAG-3′ | 892 to 874 | 812 to 892 (81 nt) |
| AS-11 | 5′-AGACCAGGAGATAATTGGAGCTGTAG-3′ | 645 to 620 | |
| AS-12 | 5′-CTCCTGGTCTGACCTCAGTGCC-3′ | 636 to 657 |
| Primer*name . | Nucleotide sequence† . | Nucleotide position‡ . | PCR-product . |
|---|---|---|---|
| cDNA | |||
| SP-1 | 5′-GAGCCAGAGGAAGAGATAGCC-3′ | −21 to −1 | |
| AS-2 | 5′-GAAAAACAGCAGTTTGCTGAGACC-3′ | 1260 to 1237 | |
| AS-3 | 5′-GGAAGTTAGCTGGAGCAGTCAGGC-3′ | 1234 to 1211 | −21 to 1234 |
| genomic-DNA | |||
| SP-4 | 5′-ctgtgtctcttgccccacaa-3′ | 20 to 1 up Ex6 | |
| AS-5 | 5′-caactcccattccctccttctgcc-3′ | 72 to 49 down Ex6 | Int5-Int6 (221 nt) |
| SP-6 | 5′-GGCGTTGTCAATGTACCTTTCAGCC-3′ | 546 to 570 | |
| SP-7 | 5′-TCCTGGTCTGACCTCAGTGCCCTG-3′ | 637 to 660 | |
| AS-8 | 5′-tgcgtatcttgcagagcc-3′ | 49 to 32 down Ex7 | Ex7-Int7 (76 nt) |
| SP-9 | 5′-GACTCAGTCTTTCAGCCCC-3′ | 812 to 830 | |
| AS-10 | 5′-CCATTGCAATGCAGGCCAG-3′ | 892 to 874 | 812 to 892 (81 nt) |
| AS-11 | 5′-AGACCAGGAGATAATTGGAGCTGTAG-3′ | 645 to 620 | |
| AS-12 | 5′-CTCCTGGTCTGACCTCAGTGCC-3′ | 636 to 657 |
SP and AS for sense and antisense primer, respectively.
The nucleotide of the AS-11 primer encoding the N211I substitution is underlined.
Position +1 refers to the first nucleotide of the initiation codon (GenBank accession number Y19039). Up and down for upstream and downstream from the indicated exon (Ex), respectively.
Int indicates intron; nt, nucleotide.