b'Sample preparationBenefits and underlying principles ofcommon nucleic acid isolation methodsOrganic extraction: Phenol-chloroform solution Benefits:(e.g., Invitrogen DNAzol and TRIzol reagents) Efficient lysis of cells and tissueAfter homogenizing the sample with TRIzol Reagent, chloroform is added, and the mixture separatesRapid denaturation of nucleases into a clear upper aqueous layer containing RNA, an interphase layer, and a pink lower organic layerStabilization of nucleic acidscontaining the DNA and protein. RNA is precipitated from the upper aqueous layer with isopropanol. DNA is precipitated from the interphase and organic layers with ethanol. Protein is precipitated from theGreat for fatty and phenolethanol supernatant with isopropanol.cartilaginous samplesSpin columns: Glass fiber, derivatized silica, or ion exchange membrane in columnBenefits:(e.g., Thermo Scientific GeneJET and Invitrogen PureLink kits) ConvenienceSamples are lysed and passed through the membrane using centrifugal or vacuum force. Wash andEase of useelution solutions are subsequently passed through the membrane, and the sample is collected into aThroughput flexibilitytube by centrifugation.Specialized equipment not requiredMagnetic beads: 0.51.0 m particles with a paramagnetic core and modified shellBenefits:(e.g., Applied Biosystems MagMAX kits and Invitrogen Dynabeads magnetic beads) No risk of cloggingIncreased target capture efficiency Samples are lysed in solution and allowed to bind nucleic acid to magnetic particles based on specific surface modifications. Application of an external magnetic field rapidly collects the particles. RoundsRapid collection andconcentration of sampleof release, washes, and recapture enable purification of the desired nucleic acid.Specialized equipment not requiredScalabilityLearn more at thermofisher.com/sampleprep 8 Contents'