Purelink hipure plasmid fp maxiprep kit

Manufactured by Thermo Fisher Scientific

The PureLink HiPure Plasmid FP Maxiprep Kit is a laboratory product designed for the rapid and efficient purification of high-quality plasmid DNA from bacterial cultures. The kit utilizes a proprietary silica-based resin to capture and purify plasmid DNA, providing a simple and reliable method for obtaining high-yield plasmid preparations.

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Lab products found in correlation

Pcr8 gw topo, by Thermo Fisher Scientific (1 mentions) Gateway lr clonase 2 enzyme mix, by Thermo Fisher Scientific (1 mentions) Protein a resin, by GenScript (1 mentions)

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2 protocols using purelink hipure plasmid fp maxiprep kit

Cloning and Characterization of Plant Promoters

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Promoter fragments of 2,009 bp (EgrNAC61 promoter; Eucgr. E01053), 3,660 bp (EgrNAC26 promoter; Eucgr. A02887) and 1,115 bp (VND6 promoter; AT5G62380) directly upstream of the translational start site were PCR-amplified from genomic DNA (Table S1), cloned into pCR8/GW/TOPO® (EgrNAC61) or pENTR/D-TOPO® (EgrNAC26 and VND6) entry vectors (Invitrogen, Carlsbad, CA) and transferred to pBGWFS7^{85 (link)} using the Gateway LR ClonaseII Enzyme Mix (Invitrogen). For Populus GUS constructs, the promoters were transferred to pMDC162^{86 (link)}. Developing secondary xylem RT-PCR products of EgrNAC26 and EgrNAC61 coding regions were cloned into pCR8/GW/TOPO® and transferred to the pUC-35S-Rfa-35S-GFP protoplast expression vector^{71 (link)} and pDEST-GBKT7 yeast expression vector^{87 (link)} similarly using LR recombination. Empty vector controls of the expression vectors were produced through LR recombination with a self-ligated pCR8/GW/TOPO® vector. Large-scale plasmid isolations for protoplast transfections were performed using the PureLink HiPure Plasmid FP Maxiprep Kit (Invitrogen), or caesium chloride purification^{88 (link)}. The pHBT::sGFP(S65T)-NOS vector^{89 (link)} was used as a fluorescent marker to assess the effect of different plasmid purification methods on protoplast transfection efficiency.

Laubscher M., Brown K., Tonfack L.B., Myburg A.A., Mizrachi E, & Hussey S.G. (2018). Temporal analysis of Arabidopsis genes activated by Eucalyptus grandis NAC transcription factors associated with xylem fibre and vessel development. Scientific Reports, 8, 10983.

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Plasmid Purification and Protein Expression

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DNA for transfection was obtained using a PureLink HiPure Plasmid FP Maxiprep Kit (Invitrogen). Proteins were obtained through transient transfection of HEK293F cells with DNA and 2 mg of polyethyleneimine per liter of culture per manufacturer’s protocol. Supernatants were harvested and sterile filtered with a 0.2 µm filter one week after transfection. All antibodies were purified using Protein A resin (Genscript) per manufacturer’s protocol. Proteins are all buffer exchanged and stored in 1× PBS.

Chen T.F., Sazinsky S.L., Houde D., DiLillo D.J., Bird J., Li K.K., Cheng G.T., Qiu H., Engen J.R., Ravetch J.V, & Wittrup K.D. (2017). Engineering Aglycosylated IgG Variants with Wild-type or Improved Binding Affinity to Human Fc gamma RIIA and Fc gamma RIIIAs. Journal of molecular biology, 429(16), 2528-2541.

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