• WorkingCellBank(ContributedbyNanciDonacki)Providesdetailedprotocolforestablishingaworkingcellbank
• MasterCellBank(ContributedbyNanciDonacki)Providesdetailedprotocolforestablishingamastercellbank
• FreezerandCellLineDatabase(contributedbyNanciDonacki)VeryusefulMicrosoftAccessdatabaseprogram(486kb)formanagingyourcelllinesstoredinfreezer.It"seasytouseandhasaverynicescreen.IthasfeatureslikeAdd,Edit,Searchandmore.(Todownloadit,forWindowsexplorer,clickthefilename;forNavigator,rightclicktosavethelink,forMac,clickandpressonthefilenameuntilmenupopsupandselecttodownload.)
• StorageofCellLine(HancockLab)(AccessIBLeonlybyIE)Preserveyourcellin10%DMSO.
• RoutineSplittingandfreezingofcells(BotwellLab)
• ThawingCellsfromLiquidNitrogen(HancockLab)(AccessibleonlybyIE)

·CryopreservationOfMammalianCellsCryopreservationOfMammalianCells(LTI)CryopreservationOfMammalianCellsCryopreservationOfMammalianCellsMammaliancellsarecryopreservedtoavoidlossbycontamination,tominimizegeneticchangeincontinuouscelllines,andto...

·CryopreservationofMammalianCells(NUNC)Mostmammaliancellscanbestoredattemperaturesbelow–130°Cformanyyears*.TheviABIlityofthecellsaftercryopreservationdependsontheirabilitytocopewiththeusr/localietyofstressesimposedonthemduringthefreezingandthawingprocedures.TheprotocolsgiveninthisTechNotewillbesuitableformostcells.Minoradjustmentsarealwayscelllinedependentandthereforeineachcasemustbefoundemperically.

·FreezingcellsinliquidN₂forstorage(AntisenseResearchGroup)

·Cell&PhageStorage(GimilaLab)

·RevivingcellsfrozeninliquidN₂(AntisenseResearchGroup)

·FreezingCellandThawingCell(Hannun/ObeidLab)

·FreezingandThawingcells(SeftonLab)

·ReactivatingFrozenCellLinesandCellGrowthforDNAPreparation(Donis-Kellerlab)Celllinesarereactivatedandgrowntoacountof1x108cells.Thecellsarepelletedandstoredfrozenat-80degreesCpriortoDNAextraction.

·PreparationofLymphocyteCellPelletforStorage(Donis-Kellerlab)Followingpropagationto1X108cells,lymphoblastoidcellsareconvenientlystoredat-80degreesCtopreservethehighmolecularweightDNAinthecellsuntiltheDNAispurified.Thisproceduredescribesthestepsrequiredtoharvestandfreezethecellsforlongtermstorage.