PSP Spin Stool DNA Kit - STRATEC Biomedical AG

More documents

Recommendations

Info

Scheme of the PSP ® Spin Stool DNA Plus KitPlease read protocols prior the start of the preparation carefullycollect a spoon of the stool sampletransfer stool sample into the Stool Collection Tube, close the tube.mix for a short time by shakingtransfer 1.4 ml of the stabilized stool sample (Stool DNA Stabilizer with stoolspecimen) into a 2.0 ml Safe-Lock Tube.for enrichment of host DNA:incubate 10 min at RT under shakingfor enrichment of bacterial DNA:incubate 10 min at 95°C on a thermomixer under shaking,add 5 Zirconia Beads II to the homogenate and vortex for 2 minspin down at 11.000 x g (11.000 rpm) for 1 mintransfer the supernatant to the InviAdsorb-Tubemix it by vortexing for 15 sec.incubate 1 min at RTspin down for 3 min at full speed.transfer the supernatant in a new 1.5 ml Receiver Tubecentrifuge the sample again at full speed for 3 min.add 25 µl Proteinase K in a new 1.5 ml Receiver Tubetransfer 800 µl of the sample supernatant to the same tubemix shortly by vortexingincubate for 10 min at 70 °C while continuously shaking on a thermomixer at900 rpm.add 400 µl Binding Buffer A (follow preparing instructions) to the lysatemix shortly by vortexing or pipetting up and downtransfer the whole mixture to the RTA Spin Filterincubate for 1 min at RTcentrifuge at 11.000 x g (11.000 rpm) for 2 min.discard the filtrate and the RTA Receiver Tubetransfer the RTA Spin Filter in a new RTA Receiver Tubepipet 500 µl Wash Buffer I onto the RTA-Spin Filtercentrifuge at 11.000 x g (11.000 rpm) for 1 mindiscard the flow-through and the RTA Receiver Tubeput the RTA Spin Filter in a new RTA Receiver Tubepipet 700 µl Wash Buffer II onto the RTA Spin Filtercentrifuge at 11.000 x g (11.000 rpm) for 1 mindiscard the flow-through and reuse the RTA Receiver Tubeto eliminate any traces of ethanol, centrifuge again for 4 min at maximum speed,discard the RTA Receiver Tubetransfer the RTA Spin Filter into a new 1.5 ml Receiver Tubepipet 100-200 µl of Elution Buffer (preheated to 70°C) directly onto the center ofthe membrane of the RTA Spin Filterincubate for 1 min at RTcentrifuge at 11.000 x g (11.000 rpm) for 1 min; discard the RTA-Spin Filter;place the eluted total DNA immediately in a refrigerator or store it at –20°CPSP ® Spin Stool DNA Kit 0213PSP ® Spin Stool DNA Plus Kit 0213
InstructionsThe following notes are valid for all protocols:Note:The centrifugation steps were made with the Centrifuge 5415 D from Eppendorf.The indicated rpm amounts are referring to this centrifuge.Protocol 1:Collection of the stool sample and stabilizationPlease read protocols prior the start of the preparation and complete preparing steps!!Note: The Stool Collection Tubes contains 8 ml of Stool DNA Stabilizer. That is a new developedbuffer formulation which enables the prelysis and stabilization of the DNA for at least 3 days atambient temperature. The Stool DNA Stabilizer is very successful even if bacterial pathogensshould be detected, which are difficult to lyse because of the structure of their cell walls.1. Open the Stool Collection Tube and collect a spoon of the stool sample.2. Transfer the spoon with the stool sample back into the Stool Collection Tube and closethe tube very tight.3. Mix the sample for a short time by shaking. That will lead to homogenization of the stoolsample.Important Notes:The collected sample can be stored at ambient temperature for at least 3 days.The storage under Stool DNA Stabilizer will lead to a better yield of bacterialpathogens with difficult to lyse cell walls. Storage time has no influence on thequality or the amount of host cell DNA.The collected sample can also be used immediately after collection for theisolation of DNA.The collected sample can be frozen at –20°C immediately after collection orafter storage at ambient temperature for a later use (for example for a secondDNA isolation).PSP ® 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: 5. Binding of the DNAAdd 200 µl of
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 and 32: insufficient mixing with Lysis Buff
Page 33 and 34: Ordering informationProduct Package

PSP Spin Stool DNA Kit - STRATEC Biomedical AG

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?