Haynes:MamCultureMedia

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(Transgenic Cell Selection)
Current revision (16:27, 2 July 2013) (view source)
(Transgenic Cell Selection)
 
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| '''Cell Type''' || '''Genotype''' || '''Transfected Plasmid''' || '''Medium''' || '''Supplements<br>(required)''' || '''Selection<br>antibiotic(s)'''
| '''Cell Type''' || '''Genotype''' || '''Transfected Plasmid''' || '''Medium''' || '''Supplements<br>(required)''' || '''Selection<br>antibiotic(s)'''
|-
|-
-
| U2OS (adherent) || Flp-in T-REx || pcDNA5/FRT, or V0200 || McCoy's 5A || 10% FBS, 1 % pen/strep || 15 ug/mL blasticidin, 200 μg/mL hygromycon
+
| U2OS (adherent) || Flp-in T-REx || pcDNA5/FRT, or V0200 || McCoy's 5A || 10% FBS, 1 % pen/strep || 15 ug/mL blasticidin, 200 μg/mL hygromycin
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Contents

Mammalian Culture Media

"Wild type" stock cells (non-transgenic)

Cell line Incomplete media Supplements for complete media (required)
HEK293 (adherent) DMEM, high glucose 10% FBS, 1% pen/strep
K-562 (suspension) Iscove's Modified Dulbecco's Medium (IMDM) 10% FBS, 1% pen/strep
SK-N-SH (adherent) Eagle's Minimum Essential Medium (EMEM) 10% FBS, 1% pen/strep
U2OS (adherent) McCoy's 5A 10% FBS, 1% pen/strep


Transgenic Cell Selection

Cell Type Genotype Transfected Plasmid Medium Supplements
(required)
Selection
antibiotic(s)
U2OS (adherent) Flp-in T-REx pcDNA5/FRT, or V0200 McCoy's 5A 10% FBS, 1 % pen/strep 15 ug/mL blasticidin, 200 μg/mL hygromycin


Transgenic Cell Maintenance

Cell Type Genotype Medium Supplements
(required)
Maintenance
antibiotic(s)
U2OS (adherent) Flp-in T-REx, untransfected McCoy's 5A 10% FBS, 1 % pen/strep 100 ug/mL Zeocin, 15 ug/mL blasticidin
U2OS (adherent) Flp-in T-REx, hygro-transfected McCoy's 5A 10% FBS, 1 % pen/strep 15 ug/mL blasticidin, 60 μg/mL hygromycin
HEK 293 (adherent) Flp-in T-REx DMEM, high glucose 10% FBS, 1 % pen/strep 100 ug/mL Zeocin, 15 ug/mL blasticidin
HEK 293 (adherent) luc Gal4-EED "23;4;9" (K. Hansen) DMEM, high glucose 10% FBS, 1 % pen/strep 0.5 ug/mL puromycin


Notes

  • FBS = fetal bovine serum
  • FBS-tf = tetracycline-free FBS (required to maintain the "off" state of dox-inducible transgenes)
  • pen/strep = penicillin and streptomycin anti fungal supplement
  • Filter sterilize all media before use
  • Store all media at 4°C


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