Phh-grd and Phh-lcch3 Cloning and Transcription

The full-length Phh-grd variants and Phh-lcch3 were amplified using GoTaq polymerase (Promega) and cloned in the pTB207 expression vector using the In-Fusion HD Cloning Kit (Takara Bio Europe SAS, Saint-Germain-en-Laye, France) as described (Lamassiaude et al., 2021 (link)). Recombinant plasmids were purified using E.Z.N.A. Plasmid DNA Mini Kit (Omega Bio-Tek, Inc., Norcross, GA), and correct cloning was confirmed by sequencing (Eurofins Genomics). cRNAs were obtained from plasmids linearized by Msc1 (Thermo Fisher Scientific) using mMessage mMachine T7 transcription kit (Thermo Fisher Scientific) following the manufacturer's instructions. cRNA concentrations were measured by spectrophotometry (NanoDrop; Thermo Fisher Scientific), and integrity of cRNAs was confirmed by running 500 ng in 1% agarose gel in TAE buffer (40 mM Tris-acetate–1 mM EDTA).

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Hashim O., Charvet C.L., Toubaté B., Ahmed A.A., Lamassiaude N., Neveu C., Dimier-Poisson I., Debierre-Grockiego F, & Dupuy C. (2022). Molecular and Functional Characterization of GABA Receptor Subunits GRD and LCCH3 from Human Louse Pediculus Humanus Humanus. Molecular Pharmacology, 102(2), 116-127.

Publication 2022

GoTaq polymerase In-Fusion HD Cloning Kit E.Z.N.A. Plasmid DNA Mini Kit mMessage mMachine T7 transcription kit NanoDrop

Corresponding Organization :

Other organizations : University of Gezira, Omdurman Islamic University, Al Baha University, Université de Tours, Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement

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Variable analysis

independent variables

Phh-grd variants
Phh-lcch3

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

controls

Positive control: Not mentioned
Negative control: Not mentioned

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