RNA Cell HTS 96 Kits

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Preparing reagents and buffers1 x 96 total RNA-extractions:add 21 ml 96-100% ethanol to the bottle Binding Buffer Radd 40 ml of 96-100% ethanol to the bottle Wash Buffer R1; Mix shortly and keep the bottlealways firmly closedadd 80 ml of 96-100% ethanol to the bottle Wash Buffer R2. mix shortly and keep the bottlealways firmly closedadjust Lysis Solution S with 1/100 volume of 1 M DTT*2 x 96 total RNA-extractions:add 42 ml 96-100% ethanol to the bottle Binding Buffer Radd 80 ml of 96-100% ethanol the bottle Wash BufferR1; mix shortly and keep the bottlealways firmly closedadd 80 ml of 96-100% ethanol to each bottle Wash BufferR2; mix shortly and keep the bottle alwaysfirmly closedadjust Lysis Solution S with 1/100 volume of 1 M DTT*4 x 96 total RNA-extractions:add 84 ml 96-100% ethanol to the bottle Binding BufferRadd 80 ml of 96-100% ethanol to the bottle Wash BufferR1; mix shortly and keep the bottlealways firmly closedadd 160 ml of 96-100% ethanol to the bottle Wash Buffer R2; mix shortly and keep the bottle alwaysfirmly closedadjust Lysis Solution S with 1/100 volume of 1 M DTT*24 x 96 total RNA-extractions:add 245 ml 96-100% ethanol to the bottle Binding BufferRadd 350 ml of 96-100% ethanol to the bottle Wash Buffer R1; mix shortly and keep the bottle alwaysfirmly closedadd 800 ml of 96-100% ethanol to the bottle Wash Buffer R2; mix shortly and keep the bottle alwaysfirmly closedadjust Lysis Solution S with 1/100 volume of 1 M DTT*Harvesting cellsCells grown in a monolayer: Cells grown in a monolayer in 96-well plates can be lysed directly in thewells. Completely remove medium by pipettingCells grown in suspension: Transfer aliquots of up to 5 x 10 5 cells into a 96-well plate. Centrifuge at240 x g (1.500 rpm) for 5 min and completely remove medium by pipetting.Note:Incomplete removal of cell culture medium will inhibit partially lysis and dilute the lysate which willaffect the yield and quality of the resulting RNA.Lysis Solution SShake Lysis Solution S gently before use to homogenize the carrier! Wait a short time because offoam formation!Cell lysates stored in Lysis Solution S are stable at –80°C for several months.Adjust Lysis Solution S with 1/100 volume of 1 M DTT. Due to the instability of dissolved DTTunder oxidative conditions do not mix the whole Lysis Solution TR with DTT in case of the kitswith 50 and 250 preparations. We recommend the preparation of a volume DTT-containing LysisSolution TR shortly before carrying out the purifications adapted to the number of samples that willbe processed. Store the remaining Lysis Solution S and DTT separately in accordance to thestorage instructions (see “Storage”, page 4).It is possible to replace DTT by β-Mercaptoethanol. In that case adjust Lysis Solution TR with1/100 volume of β-Mercaptoethanol as described above.9 InviTrap ® RNA Cell HTS 96 Kit/ C and V 0810
Equipment and reagents to be supplied by userWhen working with chemicals, always wear a suitable lab coat, disposable gloves, and protectivegoggles. For more information, please consult the appropriate material safety data sheets(MSDSs).(See our web page : www.invitek.de)○○○○○○○○○○measuring cylinder (250 ml)disposable glovesmultichannel pipet with tipsreagents reservoirs for multichannel pipets96-100% ethanoloptional: DNase A (10 mg/ml)optional: centrifugeoptional: Manifoldcentrifuge: output ≥ 2.000 x g is necessary○ Sigma-Centrifuge 4-15C or centrifuge 4K15C with plate-rotor 2 x 96 or other rotors○ Eppendorf Centrifuge 5804 / 5804 R / 5810 / 5810 R with Deepwell-Plate-Rotor (A-2-DWP)DTTPossible supplier:Eppendorf AG22331Hamburg, GermanyTel.: +49 (0) 40 53801 0Fax: +49 (0) 40 53801 556E-Mail: eppendorf@eppendorf.comInternet: www.eppendorf.comSIGMA Laborzentrifugen GmbH37507 Osterode am Harz, GermanyTel.: +49-5522-5007-0Fax: +49-5522-5007-12E-Mail: info@sigma-zentrifugen.deInternet: www.sigma-zentrifugen.deManifold:Invisorb ® 96 Vacuum Manifold or Manifolds from other supplier Vacuum pumpor another vacuum source, which develop a vacuum pressure of about 200 mbar *).Possible supplier:KNF Neuberger GmbH79112 Freiburg i.Br., GermanyTel.: +49 (0) 7664 5909 0Fax: +49 (0)7664 5909-99E-Mail: info@knf.deInternet: www.knf.deVACUUBRAND GMBH+CO KG97866 Wertheim, GermanyTel. ++49 (0) 9342 808 0Fax ++49 (0) 9342 59880E-Mail: info@vacuubrand.deInternet: www.vacuubrand.deNote: *) 200 mbar = 200hPa = 150 mmHg10 InviTrap ® RNA Cell HTS 96 Kit/ C and V 0810
Page 4: Kit content of InviTrap ® RNA Cell
Page 7 and 8: Product characteristic of the InviT
Page 9: Important notesImportant points bef
Page 13 and 14: Scheme of the InviTrap ® RNA Cell
Page 15 and 16: 16. Add again 400 µl Wash Buffer R
Page 17 and 18: 11. Repeat this washing step by add
Page 19 and 20: 13. Centrifuge at 9.000 x g (6.200
Page 21 and 22: TroubleshootingProblem Cause Commen
Page 23 and 24: When measuring RNA samples, make su
Page 25 and 26: Invitek distributorsAustraliaMedica

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