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Pseudovirus Production

Pseudovirus ProductionProduction of pseudoviruses (or capsids) using 293TT cells, including purification by ultracentrifugation through an iodixanol (Optiprep) gradient
Revised ProductionAn updated pseudovirus harvest method that removes "cold" capsids containing cellular DNA fragments
Improved MaturationA protocol modification for achieving more uniform capsid maturation
BKV HarvestProtocol modifications designed to facilitate harvest of BK polyomavirus-based reporter vectors and virions
Native MCV ProductionMethods for producing and propagating Merkel cell polyomavirus virions in cell culture
PropagationUse of a self-packaging L1+L2 plasmid for infectious amplification of pseudovirus stocks
Gel FiltrationRapid purification of pseudovirus out of crude cell lysates (or Optiprep) using agarose gel filtration columns
Dye ConjugationCoupling AlexaFluor dyes to capsids
Encapsidated DNA AnalysisExtraction and analysis of packaged DNA
Codon ModificationSilent modification of genes to remove expression-inhibitory elements (MS Excel file download)
Re-Ligation of Cloned Viral GenomesReconstitution of cloned HPV genomes by intramolecular re-ligation
Modified Hirt ExtractSpin column-based method for extracting plasmid DNA from mammalian cells
293TT CellsAn HEK-293T based cell line modified to express high levels of SV40 Large T antigen
Last updated by Buck, Christopher (NIH/NCI) [E] on Dec 21, 2018