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Pfn210a ss halotag cmv neo flexi vector

Manufactured by Promega
Sourced in United States

The PFN210A SS-HaloTag CMV-neo Flexi-Vector is a laboratory tool designed for protein expression and purification. It features a HaloTag® domain and a neomycin resistance gene, allowing for the efficient expression and selection of recombinant proteins in various cell lines.

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2 protocols using pfn210a ss halotag cmv neo flexi vector

1

Comparative Analysis of OR8H1 Orthologs

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Mammalian orthologs
of human OR8H1 have been identified via NCBI (https://www.ncbi.nlm.nih.gov/kis/info/how-are-orthologs-calculated/) and OrthoDB v11 (https://www.orthodb.org/). The protein-coding regions of the human (NCBI reference sequence:
NM_001005199.1), chimpanzee (NCBI reference sequence: XM_009440400.4),
and horse (NCBI reference sequence: NM_001391760.1) OR8H1 were amplified
from genomic DNA by polymerase chain reaction (PCR) using gene-specific
primers (Table S2), ligated with T4-DNA
ligase (#M1804, Promega, Madison, USA) EcoRI/NotI
(#R6017/ #R6435, Promega, Madison, USA) into the expression plasmid
(#pFN210A SS-HaloTag CMV-neo Flexi-Vector, Promega, Madison, USA),
and verified by Sanger sequencing (Eurofins Genomics, Ebersberg, Germany)
using vector internal primers (Table S3). The chimp DNA was obtained from a saliva sample from an animal
born to the Munich Zoological Garden before 2014. The horse DNA was
obtained from a saliva sample.
In the same way, all human OR
coding regions used in the present study (reference sequences and
selected genetic variants, Tables S4 and S5) were cloned and ligated into the pFN210A expression plasmid and
were purified using the PureYield Plasmid Midiprep System (#A2495,
Promega, Madison, USA).
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2

Cloning and Sequencing of Human OR Genes

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The protein-coding region of human OR10A6 and OR2W1 (for accession numbers see Table 2) was amplified from human genomic DNA by polymerase chain reaction (PCR), using gene-specific primers (Table S2), ligated with T4-DNA ligase (#M1804, Promega, Madison, USA) either MfeI/NotI (#R3589S/ # R0189S, New England Biolabs, Ipswich, UK) or EcoRI/NotI (#R6017/ #R6435, Promega, Madison, USA) into the expression plasmid (#pFN210A SS-HaloTag® CMV-neo Flexi®-Vector, Promega, Madison, USA), and verified by Sanger sequencing (Eurofins Genomics, Ebersberg, Germany) using vector internal primers (Table S3).
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