Product | Shelf Life | Cost/Electroporationβ | Savings |
---|---|---|---|
Amaxa® Nucleofector® Kit V (VCA-1003) | 3 Months | $15.20 | – |
Ingenio® Electroporation Kit (MIR 50112) | 1 Year | $9.16 | 40% |
Ingenio® Solution Provides Comparable Efficiency on Amaxa® Nucleofector®. Cells were electroporated in parallel with an EGFP reporter vector using a Nucleofector® II Device (Amaxa®) and either the Ingenio® Electroporation Solution or the Nucleofector® Kit V (Amaxa®) according to Amaxa®’s recommended protocol for each cell line [HL-60 (T-019), Jurkat E6-1(X-001), K-562 (T-016), THP-1 (V-001), SK-N-MC (A-023), and MCF-7 (P020)]. EGFP expressing cells were identified 24 hours post-electroporation by flow cytometry and presented as a percentage of the live cell population.
Ingenio® Solution Shows Similar Results with Amaxa® Nucleofector® and Other Electroporators. Ingenio® Solution used with Gene Pulser Electroporator Yields Comparable High Efficiency to the Amaxa® Nucleofector® II System: HL-60, Jurkat E6-1, K562, THP-1 and SK-N-MC cells were electroporated with the same EGFP reporter on the same day, using either Ingenio® Electroporation Solution in the Gene Pulser XCell (Bio-Rad®) (red bars) or the Amaxa® Solution V in Amaxa® Nucleofector® II (Lonza) (light grey bars).
Ingenio® Provides Both Superior Efficiency and Cell Viability Compared to Other Solutions. Cells were electroporated in parallel with an EGFP reporter vector using either Ingenio® Electroporation Solution, PBS or the Gene Pulser Electroporation Buffer (Bio-Rad®) on the Gene Pulser Xcell™ Eukaryotic System. (A) EGFP expressing cells were identified 24 hours post-electroporation by flow cytometry and presented as a percentage of the live cell population. (B) Cells were assayed for viablility by propidium iodide staining and flow cytometry analysis. Experiments were performed in triplicate on three separate days and the data averaged.
Product | Shelf Life | Cost/Electroporationb | Savings |
---|---|---|---|
Amaxa® Nucleofector® Kit V (VCA-1003) | 3 months | $14.64 | – |
Ingenio® Electroporation Kit (MIR 50112) | 1 year | $8.68 | 41% |
a Compatible with Amaxa® Nucleofector® II device
b Based on 2014 U.S. List prices from company websites and protocol recommendations (25 RXN kit).
Ingenio® Kits Deliver siRNA for Knockdown Using Different Electroporation Instruments. siRNA and plasmid DNA were co-electroporated with the indicated cell lines with the Ingenio® electroporation solution in 0.2 cm cuvettes using either the Gene Pulser (Bio-Rad®) or the Amaxa® Nucleofector® II (Lonza). 10µg/ml of plasmid encoding Firefly luciferase was co-electroporated with 250nM of either non-targeting siRNA control (Dharmacon Non-Targeting siRNA #1) or GL3 siRNA into Jurkat E6-1, HeLa and CHO-K1 cells. Twenty-four hours post electroporation, cells were harvested and assayed for luciferase activity. Data from independent experiments performed on different days were averaged then scaled to non-targeting siRNA control and are represented as a percentage of the control.
Pulse conditions used:
(1) Amaxa® Nucleofector® program:
a. Jurkat E6-1: X-001 with 10 x 106 cells/ml
b. HeLa: I-013 with 3 x 106 cells/ml
c. CHO-K1: U-023 with 5 x 106 cells/ml
(2) Bio-Rad® Gene Pulser Exponential Decay setting:
a. Jurkat: 150V, 950µF with 10 x 106 cells/ml
b. HeLa : 130V, 950µF with 3 x 106 cells/ml
c. CHO-K1: 150V, 900µF with 5 x 106 cells/ml
CRISPR RNP Delivery with Ingenio® Electroporation Solution. K562 and Jurkat cells were electroporated with a Cas9 protein/gRNA, ribonucleoprotein (RNP) complex, composed of 750 nM Cas9 protein (EnGen® Cas9 NLS, NEB) and 1500 nM pre-complexed two-part gRNA (IDT) targeting PPIB using the Ingenio® Electroporation Solution and a Gene Pulser Xcell™ Eukaryotic System. Exponential pulse conditions of 130V, 950 µF for K562 and 150V, 950 µF for Jurkat cells were applied to triplicate 0.2 cm cuvettes, 100 µl volume, 10 x 106 cells/ml +/- RNP complex. A T7E1 mismatch assay was used to measure cleavage efficiency at 48 hours post-transfection. Non-specific bands (NSP) were observed in the negative control of both cell lines. Cleavage efficiency was calculated based on the ratio of cleaved band intensities to the sum of cleaved and uncleaved band intensities minus the average signal of the non-specific band(s) in negative control lanes.
High Efficiency Plasmid DNA Electroporation of Human Induced Pluripotent Stem (iPS) Cells Using Ingenio®. The Ingenio® Electroporation Kit was used to transfect 2 x 106 iPS cells on the Amaxa® Nucleofector® II Device. Cells were electroporated with 8 µg ZsGreen expressing plasmid (Clontech) in 100 µl and plated in 6-well plates at 0.33 x 106 cells/well. Cells were visualized 24 hours post-transfection and
Mirus Bio offers a universal solution for your existing electroporator as well as an innovative electroporation device
Ingenio® Electroporation Solution is compatible with multiple cell types and devices.
The Ingenio® EZporator® Electroporation System is an easy-to-use electroporation system for mammalian cells.
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