The protein-coding
region
of human OR5K1 (NM_001004736.3) was derived from our previously published
OR library.99 (link) Amplification was carried
out in a touchdown approach using gene-specific primers (Table S2 ): an initial denaturation (98 °C,
3 min) and ten cycles consisting of denaturation (98 °C, 30 s),
annealing (60 °C, decreasing 1 °C per cycle down to 50 °C,
30 s), and extension (72 °C, 1 min), followed by 25 cycles of
denaturation (98 °C, 30 s), annealing (50 °C, 30 s), and
extension (72 °C, 1 min), finishing with a final extension step
in the end (72 °C, 7 min). Insertion of nucleotides into expression
vectors was done with T4-DNA ligase (#M1804, Promega, Madison, USA)
via EcoRI/NotI (#R6017/#R6435, Promega, Madison,
USA) into the expression plasmid pFN210A100 (link) and verified by Sanger sequencing using internal primers (Table S3 ) (Eurofins Genomics, Ebersberg, Germany).
region
of human OR5K1 (NM_001004736.3) was derived from our previously published
OR library.99 (link) Amplification was carried
out in a touchdown approach using gene-specific primers (
3 min) and ten cycles consisting of denaturation (98 °C, 30 s),
annealing (60 °C, decreasing 1 °C per cycle down to 50 °C,
30 s), and extension (72 °C, 1 min), followed by 25 cycles of
denaturation (98 °C, 30 s), annealing (50 °C, 30 s), and
extension (72 °C, 1 min), finishing with a final extension step
in the end (72 °C, 7 min). Insertion of nucleotides into expression
vectors was done with T4-DNA ligase (#M1804, Promega, Madison, USA)
via EcoRI/NotI (#R6017/#R6435, Promega, Madison,
USA) into the expression plasmid pFN210A100 (link) and verified by Sanger sequencing using internal primers (
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