PSP Spin Stool DNA Kit - STRATEC Biomedical AG

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AppendixGeneral notes on handling DNANature of DNAThe length and delicate physical nature of DNA require careful handling to avoid damage dueto shearing and enzymatic degradation. Other conditions that affect the integrity and stabilityof DNA include acidic and alkaline environments, high temperature, and UV irradiation.Careful isolation and handling of high molecular weight DNA is necessary to ensure it willfunction well in various downstream applications. Damaged DNA could perform poorly inapplications such as genomic Southern blotting, long-template PCR, and construction ofcosmid libraries.Handling fresh and stored material before the extraction of DNAFor the isolation of genomic DNA from cells or tissues, use either fresh samples or samplesthat have been quickly frozen in liquid nitrogen and stored at -70°C. This procedure minimizesdegradation of crude DNA by limiting the activity of endogenous nucleases.Storage of DNAStore genomic DNA at +2 to +8°C. Storing genomic DNA at -15 to -25°C can cause shearingof DNA, particularly if the DNA is exposed to repeated freeze-thaw cycles. Plasmid DNA andother small circular DNAs can be stored at +2 to +8°C or at -15 to -25°C.Drying, dissolving and pipetting DNAAvoid overdrying genomic DNA after ethanol precipitation. It is better to let it air dry than touse a vacuum, although vacuum drying can be used with caution. Plasmid DNA and othersmall circular DNAs can be vacuum-dried.To help dissolve the DNA, carefully invert the tubes several times after adding buffer and tapthe tube gently on the side. Alternatively let the DNA stand in buffer overnight at +2 to +8°C.Minimize vortexing of genomic DNA since this can cause shearing.Avoid vigorous pipetting. Pipetting genomic DNA through small tip openings causes shearingor nicking. One way to decrease shearing of genomic DNA is to use special tips that havewide openings designed for pipetting genomic DNA. Regular pipette tips pose no problem forplasmid DNA and other small fragments.Determination of concentration, yield, and purity of DNADetermination of concentration, yield, and purityDNA yields are determined from the concentration of DNA in the eluate, measured byabsorbance at 260 nm. Purity is determined by calculating the ratio of absorbance at 260 nmto absorbance at 280 nm. Pure DNA has an A260/A280 ratio of 1.7–2.1. Absorbance readingsat 260 nm should lie between 0.1 and 1.0 to be accurate. Sample dilution should be adjustedaccordingly. Use elution buffer or water (as appropriate) to dilute samples and to calibrate thespectrophotometer. Measure the absorbance at 260 and 280 nm, or scan absorbance from220–320 nm (a scan will show if there are other factors affecting absorbance at 260 nm). BothDNA and RNA are measured with a spectrophotometerPSP ® Spin Stool DNA Kit 0213PSP ® Spin Stool DNA Plus Kit 0213
Ordering informationProduct Package size Catalogue No.PSP ® Spin Stool DNA Kit 5 extractions 1038100100PSP ® Spin Stool DNA Kit 50 extractions 1038100200PSP ® Spin Stool DNA Kit 250 extractions 1038100300Single componentsLysis Buffer P 30 ml 1038101200Binding Buffer A 30 ml 1038102800InviAdsorb for 10 purifications 1038107600Wash Buffer I 30 ml 1038103300Wash Buffer II 18 ml 1038103400Elution Buffer 15 ml 1038104000Product Package size Catalogue No.PSP ® Spin Stool DNA Plus Kit 5 purifications 1038110100PSP ® Spin Stool DNA Plus Kit 50 purifications 1038110200PSP ® Spin Stool DNA Plus Kit 250 purifications 1038110300Single componentsStool DNA Stabilizer 250 ml 1038111100Stool Collection Tubes 5 tubes 1038111200(with Stool DNA Stabilizer)Binding Buffer A 30 ml 1038112800Wash Buffer I 30 ml 1038113300Wash Buffer II 18 ml 1038113400Elution Buffer 15 ml 1038114000Related productsInviMag ® Stool DNA Mini Kit 10 purifications 1438100900InviMag ® Stool DNA Mini Kit 50 purifications 1438100200InviMag ® Stool DNA Mini Kit 250 purifications 1438100300InviMag ® Stool DNA Mini Kit/ KFmL 1 x 15 purifications 2438110100InviMag ® Stool DNA Mini Kit/ KFmL 1 x 75 purifications 2438110200InviMag ® Stool DNA Mini Kit/ KF96 1 x 96 purifications 7438300100InviMag ® Stool DNA Mini Kit/ KF96 5 x 96 purifications 7438300200InviTaq Hot Start DNA Polymerase 500 Units 3020110300InviTaq Hot Start DNA Polymerase 1000 Units 3020110400Possible suppliers for Isopropanol:Carl Roth2-PropanolRotipuran >99.7%, p.a., ACS, ISOOrder no. 6752Applichem2-Propanol für die MolekularbiologieOrder no. A3928Sigma2-PropanolOrder no. 59304-1L-FPSP ® Spin Stool DNA Kit 0213PSP ® Spin Stool DNA Plus Kit 0213
Page 8 and 9: Intended useThe PSP ® Spin Stool D
Page 10 and 11: Product characteristic of the PSP
Page 13 and 14: ProcedureLysisStool samples are lys
Page 15 and 16: 5 total DNA extractions:add 250 µl
Page 17 and 18: Scheme of the PSP ® Spin Stool DNA
Page 19 and 20: 3. Second sample cleanupTransfer th
Page 21 and 22: 5. Binding of the DNAAdd 200 µl of
Page 23 and 24: InstructionsThe following notes are
Page 25 and 26: 5. Binding of the DNAAdd 400 µl of
Page 27 and 28: 6. Washing StepsPut the RTA Spin Fi
Page 29 and 30: TroubleshootingProblemComments and
Page 31: insufficient mixing with Lysis Buff

PSP Spin Stool DNA Kit - STRATEC Biomedical AG

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