Plasmid transfection in SH-SY5Y cells: best reagent?

More Frank Crittenden's questions See All

When should I add antibiotics to cell culture media?

To be clear: I'm asking about pen/strep to prevent contamination, NOT selective antibiotics. Should I add the antibiotics when I am making the media? Or should I add the antibiotics directly to...

10 November 2019 8,072 3 View

When/how should I select cells with puromycin after plasmid delivery?

I am delivering the pSpCas9(BB)-2A-puro plasmid to SH-SY5Y cells via electroporation, and I am trying to come up with a protocol for the puromycin selection. All of the published literature I can...

05 June 2019 5,146 4 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

Where should the ARS sequence be introduced on a plasmid?

Hi all, I need to introduce an ARS (autonomously replicating sequence) in my plasmid but I'm not sure which position would be the best. Does anyone have any suggestion? A picture of the plasmid...

05 August 2024 1,573 4 View

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...

05 August 2024 9,059 3 View

Why my colony PCR results of my recombinant bacterial not showing any results?

I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....

05 August 2024 2,570 3 View

Transfection in HEK293T cells?

Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...

31 July 2024 9,892 4 View

Issues with Protein Expression in Transfected cells?

Hello I am trying to create a stable cell line in HEK293 via Lipofectamine 3000 transfection. My plasmid is a CD63-IL10-GFP construct with Puromycin resistance. I am successful with the...

30 July 2024 6,648 1 View

Does anyone have experience electroporating SKOV3 cells with a large plasmid using Neon transfection?

I have been trying to electroporate SKOV3 cells with a large plasmid (11kb) without much success. Any tips?

29 July 2024 3,229 1 View

Why cannot i find my protein on cell surface after antibiotic selection of expressing plasmid?

I cannot confirm cell surface protein expression by flow cytometry even after transfection and antibiotic selection of cells. Does it take long for proteins to express on the cell surface? the...

28 July 2024 3,178 3 View

Are these cassettes suitable for expressing PETase mutant in E. coli?

I created two potential gene expression cassettes (constitutive and inducible) for expression of a mutant PETase gene on PeptiCloud using the version tree feature, which allows users to create...

28 July 2024 7,559 1 View

Can anyone help me explain my problem?

Hi, Now I am doing a single-base editing by using HiFi Cas 9, guideRNA, single stranded DNA, HDR Enhancer V2 and my tranfection mehtod is lipofection (RNAiMax). Unfortunately, after gene editing,...

26 July 2024 2,136 0 View

Reza Hadavi

you can use from:

- jetPEI®DNA transfection

https://www.ozyme.fr/gammes/pol/pdf/pol_jetprime.pdf

X-tremeGENE 9 DNA Transfection Reagent

good luck

Karen A. Darbinyan

Hello, see links below may be useful for you.

Good luck.

https://altogen.com/product/altofect-transfection-reagent/

http://m.jbc.org/content/286/6/4150.full

Jian Chen

Electroporation will be very easy for this cell line. If you are tying to electroporate cas9 related plasmid, I'll recommend that you switch to RNP. Many cas9 plasmid don't work after electroporation. Also many lentiviral or retroviral vectors don't work well after electroporation. If you need electroporation, Celetrix electroporation will make your transfection simple and easy.