1st primer for C-term tagged
As shown below, insert a part of gene-specific sequence and prepare 1st PCR primers for C-terminal tags (1st_ORF_CF and 1st_ORF_CF 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.
Tm value calculation of forward primer and reverse primer can be done with the 1st primer design tool in the download section.
1st PCR Primer_C (10 μM each)
| Primer name | Primer sequence 5′ to 3′ |
| 1st_ORF_CF | CACAAAACAT TTCCCTACAT ACAACTTTCA ACTTCCTATT + ORFATG NNNNNNNNNNNNNNNNNN (N=18) |
| 1st_ORF_CR | AGTACCTCCC TGCTGGAGACC + ORFRRRRRRRRRRRRRRRRRR (R=18) |