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Table 1.

Primer sequences

PrimerSequence
Degenerate primers  
    HATP-for-1 Clayborne 5′GRT GNG TDA TRT CRT CRT NNG G-3 
    HATP-rev-1 Clayborne 5′GGN GTN AAY ATG GAR CAN GC-3′ 
RACE PCR primers  
    GeneRacer™ 5′Primer 5′-CGA CTGG AGC ACG AGG ACA CTG A-3′ 
    GeneRacer™ 3′Primer 5′-GCT GTC AAC GAT ACG CTA CGT AAC G-3′ 
    Zf vatB1-5′Race 5′-ACG GGC AGC AGA CAC CTC TCT C-3′ 
    Zf vatB1-3′Race 5′-CTG AAC CTG GCC AAC GAT CCT ACC-3′ 
    Zf vatB2-5′Race 5′GTA AGG CCT CGG CAT AAG AGC TCA T-3′ 
    Zf vatB2-3′Race 5′GGC CAA TGA TCC CAC GAT TGA A-3′ 
Sequencing primers  
    M 13 forward 5′CTGGCCGTCGTTTTAC-3′ 
    M 13 reverse 5′CAGGAAACAGCTATGA-3′ 
    T7 forward 5′-TAATACGACTCACTATAGGG-3′ 
    V5C-term reverse 5′-ACCGAGGAGAGGGTTAGGGAT-3′ 
    ZF 3′vatB1 vorne 5′GAT GAC TTG CTG TAC CTG-3′ 
    Zf 3′vatB1 hinten 5′TAT TTC CCT TTG AGA TGT-3′ 
    Zf 3′Race vatB2 vorne 5′AAA GAT GTC CAG GCA ATG-3′ 
    Zf 3′Race vatB2 hinten 5′TGT AAA CTC TGT CTG GCC-3′ 
Amplification of ORFs  
    Zf vatB1 99/22 5′-ATG TCG ACG TTA GTA GCG AAC C-3′ 
    Zf vatB1 1595/22 rc 5′-AGA CTC TCT CGG GTA GAA TTC G-3′ 
    Zf vatB2 149/23 5′-ATG AAG GCT CTT AGG GGA ATG GT-3′ 
    Zf vatB2 1648/22 rc 5′-AGC CAG AGT GCT CTG AGG AAT C-3′ 
Reverse transcriptase PCR  
    Zf vatB1 123/18 5′-CCC GTG GAT CTA AAT GGC-3′ 
    Zf vatB1 576/23 rc 5′-TCT CCT CTG GGT AGA TAC GAC AC-3′ 
    Zf vatB2 172/24 5′-GAG CGG AGC CGT GAG CGA AAT ATC-3′ 
    Zf vatB2 580/24 rc 5′-CAC CGC AGG TCC TCG ATC AAT TGG-3′ 
    Zf β2-microglobulin 963/23* 5′-ACC AAA CAC CCT GAT CTG CTA TG-3′ 
    Zf β2-microglobulin 1669/21 rc* 5′-CCC CTG CAC ACC AAG AAC AAG-3′ 
PrimerSequence
Degenerate primers  
    HATP-for-1 Clayborne 5′GRT GNG TDA TRT CRT CRT NNG G-3 
    HATP-rev-1 Clayborne 5′GGN GTN AAY ATG GAR CAN GC-3′ 
RACE PCR primers  
    GeneRacer™ 5′Primer 5′-CGA CTGG AGC ACG AGG ACA CTG A-3′ 
    GeneRacer™ 3′Primer 5′-GCT GTC AAC GAT ACG CTA CGT AAC G-3′ 
    Zf vatB1-5′Race 5′-ACG GGC AGC AGA CAC CTC TCT C-3′ 
    Zf vatB1-3′Race 5′-CTG AAC CTG GCC AAC GAT CCT ACC-3′ 
    Zf vatB2-5′Race 5′GTA AGG CCT CGG CAT AAG AGC TCA T-3′ 
    Zf vatB2-3′Race 5′GGC CAA TGA TCC CAC GAT TGA A-3′ 
Sequencing primers  
    M 13 forward 5′CTGGCCGTCGTTTTAC-3′ 
    M 13 reverse 5′CAGGAAACAGCTATGA-3′ 
    T7 forward 5′-TAATACGACTCACTATAGGG-3′ 
    V5C-term reverse 5′-ACCGAGGAGAGGGTTAGGGAT-3′ 
    ZF 3′vatB1 vorne 5′GAT GAC TTG CTG TAC CTG-3′ 
    Zf 3′vatB1 hinten 5′TAT TTC CCT TTG AGA TGT-3′ 
    Zf 3′Race vatB2 vorne 5′AAA GAT GTC CAG GCA ATG-3′ 
    Zf 3′Race vatB2 hinten 5′TGT AAA CTC TGT CTG GCC-3′ 
Amplification of ORFs  
    Zf vatB1 99/22 5′-ATG TCG ACG TTA GTA GCG AAC C-3′ 
    Zf vatB1 1595/22 rc 5′-AGA CTC TCT CGG GTA GAA TTC G-3′ 
    Zf vatB2 149/23 5′-ATG AAG GCT CTT AGG GGA ATG GT-3′ 
    Zf vatB2 1648/22 rc 5′-AGC CAG AGT GCT CTG AGG AAT C-3′ 
Reverse transcriptase PCR  
    Zf vatB1 123/18 5′-CCC GTG GAT CTA AAT GGC-3′ 
    Zf vatB1 576/23 rc 5′-TCT CCT CTG GGT AGA TAC GAC AC-3′ 
    Zf vatB2 172/24 5′-GAG CGG AGC CGT GAG CGA AAT ATC-3′ 
    Zf vatB2 580/24 rc 5′-CAC CGC AGG TCC TCG ATC AAT TGG-3′ 
    Zf β2-microglobulin 963/23* 5′-ACC AAA CAC CCT GAT CTG CTA TG-3′ 
    Zf β2-microglobulin 1669/21 rc* 5′-CCC CTG CAC ACC AAG AAC AAG-3′ 

RACE, rapid amplification of cDNA ends; ORF, open reading frame; PCR,polymerase chain reaction; Zf, zebrafish.

*

Primers were designed and named on the basis of a sequence ofβ2-microglobulin (GenBank accession number L05384; Ono et al., 1993).

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