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Griffin:Puromycin Selection - OpenWetWare

(2 day ago) The working puromycin concentration for mammalian cell lines ranges from 1-10 µg/ml. Prior to using the puromycin antibiotic, titrate the selection agent to determine the optimal concentration for target cell line. Use the lowest concentration that kills 100% of non-transfected cells in 3-5 days from the start of puromycin selection.

https://openwetware.org/wiki/Griffin:Puromycin_Selection

DA: 15 PA: 33 MOZ Rank: 48

Puromycin and Blasticidin Titration 2019

(9 day ago) Protocol: Puromycin and blasticidin titration Brief Description: A lentiviral construct carrying an antibiotic resistance gene will allow selection of the cell population that has integrated that transgene. This protocol describes a method to establish a dose of puromycin or blasticidin to select the transgenic cells. The pRosetta (aka

https://portals.broadinstitute.org/gpp/public/dir/download?dirpath=protocols/production&filename=Puromycin_and_blasticidin_titration_protocol_Feb2019.pdf

DA: 26 PA: 24 MOZ Rank: 50

A puromycin selectable cell line for the enrichment of

(18 day ago) The resulting transgenic-ES cell line (Sim1-Puro Clone 6E) was used for all subsequent studies. Increased purity of Sim1 + cells after puromycin selection. To test for PAC expression in Sim1 + cells, RW4 and Sim1-Puro EBs were subjected to induction and selection (2 − /6 + /1p) protocols as illustrated in Fig. 2a. Visual assessment of calcein

https://stemcellres.biomedcentral.com/articles/10.1186/s13287-015-0213-z

DA: 29 PA: 35 MOZ Rank: 64

When/how should I select cells with puromycin after

(15 day ago) 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.

https://www.researchgate.net/post/When_how_should_I_select_cells_with_puromycin_after_plasmid_delivery

DA: 20 PA: 50 MOZ Rank: 94

DEVELOPMENT OF STABLE TRANSFECTED CELL LINE

(7 day ago) Puromycin 2.5 mg/ml stock solution Procedure: Titrating G418, Hygromycin, and Puromycin (Kill Curves) Since each cell line has a different sensitivity to G418, hygromycin or puromycin, you should determine the optimal concentration of drug for selection. 1. Split confluent cells 1:5 in 10 ml DMEM + 10% FBS media. 2.

https://groups.molbiosci.northwestern.edu/morimoto/research/Protocols/II. Eukaryotes/C. Stable Cell Lines/2. Gen's protocol.pdf

DA: 33 PA: 50 MOZ Rank: 50

Puromycin | Data sheet | InvivoGen

(1 day ago) Dying cells detach from the plates, allowing easy and early identification of transformant clones. Suggested working conditions for selection in some mammalian cells are listed below. Cell line Medium Puromycin conc. Ref. B16 (Mouse melanocytes) RPMI 1-2 μg/ml 2, 3 HEK293 (Human kidney cells)DMEM 0.5-10 μg/ml 4, 5, 6

https://www.invivogen.com/sites/default/files/invivogen/products/files/puromycin_tds.pdf

DA: 17 PA: 50 MOZ Rank: 80

CRISPR / Cas9 KO Plasmid PROTOCOL and HDR Plasmid

(2 day ago) target cell line. Use the lowest concentration that kills 100% of non- transfected cells in 3–5 days from the start of puromycin selection. • 48–96 hours post-transfection, aspirate the medium and replace with fresh medium containing puromycin at the appropriate concentration. • Select cells for a minimum of 3–5 days.

http://datasheets.scbt.com/protocols/CRISPR_protocol.pdf

DA: 19 PA: 30 MOZ Rank: 49

Lentiviral Transduction Protocol - Sigma-Aldrich

(2 day ago) Puromycin titration (kill curve) should be performed when working with a new cell type. 1. Plate 1.6 x 104 cells into wells of a 96 -well plate with 120 µl fresh media. 2. The next day add 500 – 10,000 ng/ml of puromycin to selected wells. 3. Examine viability every 2 days.

https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/General_Information/lentiviraltransdprotocol.pdf

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Does anyone have a good protocol for puromycin selection

(3 day ago) I have also used transient selection of puromycin for generating knockouts in HeLa cells. I add 2ug/ml of puromycin containing complete medium after 24h of transfection and replace the medium next

https://www.researchgate.net/post/Does_anyone_have_a_good_protocol_for_puromycin_selection_for_pSpCas9BB-2A--Puro_transfectants

DA: 20 PA: 50 MOZ Rank: 50

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