Nucleofector sg kit

Manufactured by Lonza

The Nucleofector SG kit is a laboratory equipment product designed for the transfection of nucleic acids, such as plasmid DNA or RNA, into a variety of cell types. It provides a convenient and efficient method for introducing genetic material into cells. The core function of the Nucleofector SG kit is to facilitate the delivery of nucleic acids into cells using an electrical pulse-based technology.

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Lab products found in correlation

Nucleofector sf kit, by Lonza (2 mentions) Dual luciferase reporter assay kit, by Promega (1 mentions) Quickchange 2 site directed mutagenesis kit, by Agilent Technologies (1 mentions) Dual luciferase reporter assay, by Promega (1 mentions) Amaxa nucleofector, by Lonza (1 mentions) Pcdna3.1 c k dyk, by GenScript (1 mentions)

2 protocols using nucleofector sg kit

Functional Characterization of TNFAIP3 SLE Risk Variants

Cited 3 times

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Non-risk or risk sequences surrounding rs10499197 or rs58905141 were cloned from EBV B cells positive for the non-risk or risk TNFAIP3 SLE risk haplotype tagged by rs10499197 into a minimal promoter firefly luciferase plasmid, pGL4.23 (Promega) (Supplementary Table S2) (Wang et al., 2013 (link); Pasula et al., 2020 (link); Gopalakrishnan et al., 2022 (link)). Quick change II site directed mutagenesis kit (Agilent Technologies; #200523) was used following manufacturer’s instructions to allele swap rs9494868 (Supplementary Table S2). Empty vector, non-risk clone, or risk clone was transiently co-transfected with the transfection control renilla luciferase plasmid, pRL-TK, using a 4D Amaxa Nucleofector Unit (Lonza) for Jurkat cells (Nucleofector SE kit; #V4XC-1032), THP-1 cells (Nucleofector SG kit; #V4XC-3032), and EBV B cells (Nucleofector SF kit; #V4XC-2032). Twenty-four hours post transfection, cells were stimulated with P/I for 2 h. Enhancer activity was determined using the Promega Dual-Luciferase Reporter Assay kit (Promega) following manufacturer’s protocol. Relative luciferase units (RLU) were determined by normalizing the firefly luciferase activity to the Renilla luciferase activity. RLU were normalized to the vector-only control and reported as normalized RLU.

Pasula S., Gopalakrishnan J., Fu Y., Tessneer K.L., Wiley M.M., Pelikan R.C., Kelly J.A, & Gaffney P.M. (2022). Systemic lupus erythematosus variants modulate the function of an enhancer upstream of TNFAIP3. Frontiers in Genetics, 13, 1011965.

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Functional Validation of TNIP1 Variants

Cited 1 time

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DNA sequences, approximately 350 bp in length, surrounding selected non-risk or risk TNIP1 variants were PCR amplified (Supplementary Table 1) and cloned into a minimal promoter luciferase plasmid, pGL4.23 (Promega, Madison, WI). CREB1 (GenScript; #OHu22955D) and BHLHE40/DEC1 (GenScript; #OHu17520) cloned into flag-tag overexpression cloning vector, pcDNA3.1/C-(K)DYK were purchased from GenScript (Piscataway, NJ). Indicated plasmids was transiently co-transfected with the pRL-TK plasmid into Jurkat (Nucleofector SE kit, #V4XC-1032), THP-1 (Nucleofector SG kit, #V4XC-3032), or EBV B (Nucleofector SF kit, #V4XC-2032) cells using Amaxa Nucleofector (Lonza, Portsmouth, NH; SF kit). The pRL-TK plasmid was used for normalization and to calculate transfection efficiency. Twenty-four hours post transfection, cells were treated with P/I (50 ng/ml, 500 ng/ml) for 2 h, then the enhancer activity was measured using the Dual-Luciferase reporter assay (Promega, Madison, WI) as previously described (29 (link)).

Pasula S., Tessneer K.L., Fu Y., Gopalakrishnan J., Pelikan R.C., Kelly J.A., Wiley G.B., Wiley M.M, & Gaffney P.M. (2020). Risk Variants with Opposing Functional Effects Result in Hypomorphic Expression of TNIP1 and Other Genes within a 3D Chromatin Network. Arthritis & rheumatology (Hoboken, N.J.), 72(5), 780-790.

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