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Hygromycin selection

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Hygromycin selection is a laboratory tool used in molecular biology and genetic engineering to identify and select cells that have successfully integrated a gene of interest. It functions by introducing a hygromycin resistance gene into the target cells, allowing them to survive and grow in the presence of the antibiotic hygromycin, while cells without the resistance gene are killed.

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Lab products found in correlation

Fugene 6, by Promega (1 mentions) Phusion polymerase, by New England Biolabs (1 mentions) In fusion hd cloning system, by Takara Bio (1 mentions) Pcep4, by Thermo Fisher Scientific (1 mentions)

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Hygromycin (298 citations)

2 protocols using hygromycin selection

1

Overexpressing Telomerase in ATM-Mutant HIFs

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The catalytic subunit of human telomerase on vector pBABE-hygro-hTERT plasmid (Addgene, #1773) was packaged into retroviral particles at 32°C using the amphotropic Phoenix A cell line and Fugene-6 (Promega, Madison, WI). Wild Type and ATM-mutant HIFs were transduced and resistant colonies pooled and expanded after 15 days of hygromycin selection (Sigma Chemical, Saint Louis, MO).
Carranza D., Torres-Rusillo S., Ceballos-Pérez G., Blanco-Jimenez E., Muñoz-López M., García-Pérez J.L, & Molina I.J. (2018). Reconstitution of the Ataxia-Telangiectasia Cellular Phenotype With Lentiviral Vectors. Frontiers in Immunology, 9, 2703.
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2

Generation of TVA-G Monosynaptic Tracing Allele

Cited 1 time
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To generate the TVA-G monosynaptic tracing allele, cDNA encoding TVA and Rabies-G was excised from pBOB-synP-HTB (Addgene plasmid #30195; (Miyamichi et al., 2011 (link)) and subcloned into a custom pminiTol2 expression construct (Addgene plasmid #31829; (Balciunas et al., 2006 ) harboring a PGK promoter (pPL451; (Liu et al., 2003 (link)), hygromycin resistance cassette (pCEP4, ThermoFisher Scientific) and BGH poly-A sequence (pcdDNA3.1, Invitrogen) using high-fideity Phusion polymerase (New England Biolabs) and In-Fusion HD Cloning System (Clontech). The TVA-G Tol2 transfer vector was nucleofected into Hb9::GFP ESCs with pCMV-Tol2 transposase (Addgene plasmid #31823, (Balciunas et al., 2006 ) using Amaxa Nucleofector Kit for Neural Stem Cells (Lonza), followed by hygromycin selection (Sigma-Aldrich). Selected clones were differentiated using standard RA/SAG protocol to generate MNs. A red fluorescent protein variant (dsRed) of SADB19ΔG RABV was added at low titer to Day 6 EBs to assess efficiency of viral transfer.
Hoang P.T., Chalif J.I., Bikoff J.B., Jessell T.M., Mentis G.Z, & Wichterle H. (2018). Subtype Diversification and Synaptic Specificity of Stem Cell-Derived Spinal Interneurons. Neuron, 100(1), 135-149.e7.
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