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Nepa21 electroporator

Manufactured by Bulldog Bio
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The NEPA21 electroporator is a compact and versatile device designed for efficient electroporation of cells. It generates the electrical pulses required to temporarily permeabilize cell membranes, enabling the introduction of various molecules, such as DNA, RNA, or other compounds, into the cells. The NEPA21 provides a user-friendly interface and precise control over the electroporation parameters to optimize the process for different cell types and applications.

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Lab products found in correlation

Synthetic sgrna, by Synthego (2 mentions) 2 mm gap cuvette, by Bulldog Bio (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Femtojet 4i microinjector, by Eppendorf (1 mentions)

Spelling variants of this product

NEPA21 Type II Super Electroporator (2 citations)

4 protocols using nepa21 electroporator

1

Plasmid Transfection and siRNA Knockdown

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pcDNA3-HA-ADH (ALDH1A1) plasmid was a gift from Steven Johnson (Addgene plasmid # 11610), pcDNA3.1-C-(k)DYK-CYP26A1 was purchased from GenScript (Piscataway, NJ). ON-TARGETplus SMARTPool Human ALDH1A1 siRNA (Cat. ID: L-008722), ON-TARGETplus SMARTPool Human POLQ siRNA (L-015180-), ON-TARGETplus Human siRARA-1 (5’-GACAAGAACUGCAUCAUCA-3’), siRARA-4 (5’-GAGCAGCAGUUCUGAAGAG-3’), and non-targeting control siRNA (5′-UUCUCCGAACGUGUCACGU-3′) were purchased from Horizon (Lafayette, CO). MISSION shALDH1A1-1 (5’-GCTGATTTAATCGAAAGAGAT-3’), shALDH1A1-4 (5’-GCCAAATCATTCCTTGGAATT-3’) were purchased from Millipore Sigma (Burlington, MA). Plasmids (0.5 μg/ml) and siRNA (100 nM) were transfected into cells using either electroporation (NEPA-21 Electroporator, Bulldog Bio, Portsmouth, NH), or Lipofectamine 2000 (Thermo Fisher Scientific, Waltham, MA), according to the manufacturer’s instruction.
Lavudi K., Banerjee A., Li N., Yang Y., Cai S., Bai X., Zhang X., Li A., Wani E., Yang S.M., Zhang J., Rai G., Backes F., Patnaik S., Guo P, & Wang Q.E. (2023). ALDH1A1 promotes PARP inhibitor resistance by enhancing retinoic acid receptor-mediated DNA polymerase θ expression. NPJ Precision Oncology, 7, 66.
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2

Retina Electroporation: Ex Vivo and In Vivo

Cited 1 time
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Ex vivo and in vivo retina electroporation was carried out as previously described6 (link),18 (link),22 (link),51 (link) using the FemtoJet 4i microinjector (Eppendorf, Cat#5252000021) and NEPA21 Electroporator (Bulldog Bio). For ex vivo electroporation, 5 pulses of 25V, 50ms each with 950ms intervals were applied to dissected retinas. For in vivo electroporation, 5 pulses of 80V, 50ms each with 950ms intervals were applied to neonatal mouse pups. All ex vivo and in vivo electroporation experiments were repeated with at least three biological replicates. Plasmids were electroporated with a concentration of 500ng/ul to 1ug/ul per plasmid.
Keyt B.A., Paoni N.F., Refino C.J., Berleau L., Nguyen H., Chow A., Lai J., Peña L., Pater C, & Ogez J. (1994). A faster-acting and more potent form of tissue plasminogen activator.. Proceedings of the National Academy of Sciences of the United States of America, 91(9), 3670-3674.
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3

CRISPR-Mediated Genetic Editing in Organoids

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Confluent hiSIIN organoids were dissociated to single cells as described above and resuspended in 100 μL OPTI-MEM. Ribonucleoprotein (RNP) complexes were formed by mixing 1.64 μL (0.1 nmol) Alt-R Cas9 (IDT) with 3 μL (0.3 nmol) synthetic sgRNA (Synthego) and incubating for 10–20 minutes at room temperature. Cells were then added to the RNP mix, 100 μL transferred to a 2 mm gap cuvette (Bulldog Bio), and electroporated using a NEPA21 electroporator (Bulldog Bio) with the following poring pulse parameters: 175 V, 5 msec length, 50 msec interval, 2 pulses, 10% decay rate, + polarity; and transfer pulse parameters: 20 V, 50 msec length, 50 msec interval, 5 pulses, 40% decay rate, +/− polarity. Electroporated organoids were resuspended gently in pre-warmed minimal media and incubated at 37°C for 15 minutes before plating in Matrigel. sgRNAs used for electroporation are: H-2Kb: 5’-CAAUGAGCAGAGUUUCCGAG-3’; previously published B2m sequence79 : 5’-UUGAAUUUGAGGGGUUUCUG-3’.
Westcott P.M., Sacks N.J., Schenkel J.M., Ely Z.A., Smith O., Hauck H., Jaeger A.M., Zhang D., Backlund C.M., Beytagh M.C., Patten J., Elbashir R., Eng G., Irvine D.J., Yilmaz O.H, & Jacks T. (2021). Low neoantigen expression and poor T-cell priming underlie early immune escape in colorectal cancer. Nature cancer, 2(10), 1071-1085.
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4

CRISPR-Mediated Genetic Editing in Organoids

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Confluent hiSIIN organoids were dissociated to single cells as described above and resuspended in 100 μL OPTI-MEM. Ribonucleoprotein (RNP) complexes were formed by mixing 1.64 μL (0.1 nmol) Alt-R Cas9 (IDT) with 3 μL (0.3 nmol) synthetic sgRNA (Synthego) and incubating for 10–20 minutes at room temperature. Cells were then added to the RNP mix, 100 μL transferred to a 2 mm gap cuvette (Bulldog Bio), and electroporated using a NEPA21 electroporator (Bulldog Bio) with the following poring pulse parameters: 175 V, 5 msec length, 50 msec interval, 2 pulses, 10% decay rate, + polarity; and transfer pulse parameters: 20 V, 50 msec length, 50 msec interval, 5 pulses, 40% decay rate, +/− polarity. Electroporated organoids were resuspended gently in pre-warmed minimal media and incubated at 37°C for 15 minutes before plating in Matrigel. sgRNAs used for electroporation are: H-2Kb: 5’-CAAUGAGCAGAGUUUCCGAG-3’; previously published B2m sequence79 : 5’-UUGAAUUUGAGGGGUUUCUG-3’.
Westcott P.M., Sacks N.J., Schenkel J.M., Ely Z.A., Smith O., Hauck H., Jaeger A.M., Zhang D., Backlund C.M., Beytagh M.C., Patten J., Elbashir R., Eng G., Irvine D.J., Yilmaz O.H, & Jacks T. (2021). Low neoantigen expression and poor T-cell priming underlie early immune escape in colorectal cancer. Nature cancer, 2(10), 1071-1085.
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