1st primer for N-term tagged

1st primer for N-term tagged

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As shown below, insert a part of gene-specific sequence to prepare 1st PCR primers for N-terminal tags (1st_ORF_NF and 1st_ORF_NF below). The reverse primer inserts the 18 mer of reverse complementary strand excluding Stop codon.

Tm value calculation of forward primer and reverse primer can be done with the 1st primer design tool in the download section.

nterminus 1st PCR primer

 

1st PCR Primer_N ( 10 μM each)

Primer name  Primer sequence 5′ to 3′
1st_ORF_NF  CCAGCAGGGAGGTACT + ORFATG NNNNNNNNNNNNNNNNNN (N=18)
1st_ORF_NR  CCTTATGGCCGGATCCAAGAGCTCTTTTTTTTTTTTA + ORFRRRRRRRRRRRRRRRRRR (R=18)