For transfections, lentiviral vectors were generated using the following plasmids: (i) pMDG-VSVG envelope encoding a vesicular stomatitis virus, G glycoprotein. (ii) pCMV-dR8.91 packaging plasmid containing the (gag/pol) structural and polymerase genes. (iii) pMNDU3-OKSM-PGK-EGFP-WPRE transgene construct containing a polycistronic, 4-factor iPSC induction cassette (OCT4, KLF4, SOX2, and c-Myc) and a PGK promoter driven EGFP green fluorescent protein. For the bioreactor, 10 mg preps of plasmid DNA were grown up in chemically competent DH10B Escherichia coli obtained from Life Technologies and purified using an Endotoxin-Free Giga-Prep Kit (Qiagen). Purified plasmids had endotoxin levels ≤0.5 EU/ml.
Endotoxin free giga prep kit
Manufactured by Qiagen
Sourced in Germany
The Endotoxin-Free Giga-Prep Kit is a laboratory equipment designed for the purification of high-quality plasmid DNA. It is capable of yielding large amounts of endotoxin-free plasmid DNA from bacterial cultures.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using endotoxin free giga prep kit
1
Lentiviral Vector Generation for iPSC
2
Lipid-based Delivery of Genetic Cargo
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The lipids 1,2-distearoyl-sn-glycero-3-phosphorylcholine (DSPC) and 1,2-distearoyl-sn-glycero- . pDNA encoding TdTomato 15 was purchased from Addgene (Cambridge, MA) and prepared using a Qiagen Endotoxin-free Giga prep kit (Hilden, Germany). Minicircle DNA (mcDNA) was generated by transforming the mcDNA producer plasmids into E. coli ZYCY10P3S2T and purification of resulting mcDNA vectors using the Qiagen Endotoxin-free plasmid purification kit as previously described. 16 siRNA against firefly luciferase 17 was purchased from IDT (Coralville, IA).
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