Instruction for the Invisorb Spin Cell Mini Kit

Buccal swabs:To collect a sample, scrape the swab firmly against inside of each cheek 6 times. Air-dry theswab for at least 2h after collection or use them fresh prepared. Ensure that the personproviding the sample has not consumed any food or drink in the 30 min prior to samplecollection. Best results are obtained if the swab stays in the lysis solution during lysisprocedure. Use of poor quality starting material influences yield of purified DNA. This protocolis recommended for every common swab, like e.g. the following swab types: C:E:P: (OmniSwab from Whatman), cotton swab, Superswabs, Copan-Swab or DRACON tip fromHardwood Products company, CellProjects or Hain Diagnostika)Cells grown in suspension:Spin up to 1 x 10 7 cells for 5 min at 300 x g (1.500 rpm* ). Discard supernatant and remove allmedia completely, taking care not to disturb the cell pellet. At this point cells may be frozen (at -20°C or - 80°C) for future use or may be used immediately.Cells grown in a monolayer:Aspirate the media completely from the cells and continue immediately with the lysis step.Alternatively cells can be detached by trypsination (cultivation in larger culture vessels, likedishes > 35 mm, flasks > 12.5 cm 2 ). Transfer cells to a 50 ml reaction tube, pellet bycentrifugation at 300 x g (1.500 rpm* ) for 5 min and aspirate supernatant completely.ProcedureLysisSamples are lysed at elevated temperatures. Lysis is performed in the presence of LysisBuffer D.Binding genomic DNABy adding Binding Buffer HL to the lysate, optimal binding conditions will be adjusted.All lysate is applied to a spin filterRemoving residual contaminationsContaminants are efficiently washed away using Wash Buffer, while the genomic DNAremains bound to the spin filter.ElutionGenomic DNA is eluted from the spin filter using 30 - 200 Al Elution Buffer D. The eluted DNAis ready for use in different downstream applications. Eluted DNA stored at 4 – 8°C is stable for2 months or for more than 5 years if stored at -20°C.Yield and quality of genomic DNAThe amount of purified DNA using the Invisorb ® Spin Cell Mini Kit procedure, depends on thestrain, the number of cells in the sample, the sample source, transport conditions, storage, andage of the sample.Yield and quality of isolated genomic DNA is suitable for any molecular-diagnostic detectionsystem. The diagnostic tests should be performed according to manufacturers’ specifications.8Invisorb ® Spin Cell Mini Kit 032010