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Mag-Bind® FFPE RNA 96

Table of Contents

Introduction.....................................................................................2

Principle.............................................................................................2

Starting Materials...........................................................................2

Kit Contents......................................................................................3

Preparing Reagents/Storage..................................................4-5

Mag-Bind® FFPE RNA 96 Protocol............................................6

Mag-Bind® FFPE RNA 96 Protocol with Xylene.................10

Troubleshooting Guide..............................................................15

Manual Revision: March 2018

Содержание Mag-Bind FFPE RNA 96

Страница 1: ...Mag Bind FFPE RNA 96 Kit M2551 00 1 x 96 preps M2551 01 4 x 96 preps March 2018...

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Страница 3: ...Contents Introduction 2 Principle 2 Starting Materials 2 Kit Contents 3 Preparing Reagents Storage 4 5 Mag Bind FFPE RNA 96 Protocol 6 Mag Bind FFPE RNA 96 Protocol with Xylene 10 Troubleshooting Guid...

Страница 4: ...stion After two wash steps purified RNA is eluted with RNase free water Starting Materials Since standard formalin fixation and paraffin embedding procedures cause significant fragmentation of nucleic...

Страница 5: ...8 4 mL RML Buffer 35 mL 140 mL MFB Buffer 20 mL 80 mL GFC Buffer 10 mL 40 mL RNA Wash Buffer II 25 mL 2 x 50 mL LPA Buffer 1 1 mL 4 4 mL DNase I Digestion Buffer 2 x 5 mL 2 x 25 mL Mag Bind DNase I 1...

Страница 6: ...imize RNA degradation Wear gloves protective goggles and take great care when working with chemicals 1 Dilute RNA Wash Buffer II with 100 ethanol and store at room temperature Kit 100 Ethanol to be Ad...

Страница 7: ...should be stored at 2 8 C for long term use Proteinase K Solution can be stored at room temperature for up to 12 months For long term storage store Proteinase K Solution at 2 8 C All remaining compon...

Страница 8: ...uffer according to the Preparing Reagents section on Page 4 Set water bath or heat block to 55 C Set water bath or heat block to 80 C Set water bath or heat block to 37 C 1 Add 250 L RML Buffer into e...

Страница 9: ...en add 10 L LPA Buffer 11 Let sit at room temperature for 5 10 minutes 12 Place the plate onto a magnetic separation device for deep well plates and wait 7 10 minutes or until the Mag Bind Particles S...

Страница 10: ...0 times Note GFC Buffer must be diluted with 100 ethanol prior to use Please see instructions on Page 4 21 Let sit at room temperature for 3 5 minutes 22 Place the plate onto a magnet separation devic...

Страница 11: ...he Mag Bind Particles SC thoroughly by vortexing for 30 seconds or pipetting up and down 30 times 31 Let sit at room temperature for 10 minutes 32 Place the plate onto a magnet separation device to ma...

Страница 12: ...or 96 well plates MSD 01B or MSD 01 Water bath or heat block capable of 55 C Water bath or heat block capable of 80 C Water bath or heat block capable of 37 C Sealing film Before Starting Prepare RNA...

Страница 13: ...8 10 for a second ethanol wash step 12 Air dry the tissue pellet for 10 20 minutes Note It is critical to completely dry the sample before the Proteinase K digestion step Ethanol residue will effect t...

Страница 14: ...e transferred twice to process whole sample 21 Aspirate and discard the cleared supernatant 22 Remove the plate from the magnetic separation device 23 Add 400 L RNA Wash Buffer II Resuspend the Mag Bi...

Страница 15: ...emove the plate from the magnetic separation device 33 Add 400 L RNA Wash Buffer II Resuspend the Mag Bind Particles SC thoroughly by vortexing for 20 seconds or pipetting up and down 10 20 times 34 P...

Страница 16: ...lace the plate onto a magnet separation device to magnetize the Mag Bind Particles SC Wait 3 5 minutes or until all the Mag Bind Particles SC are cleared from solution 41 Transfer the cleared supernat...

Страница 17: ...not prepared correctly Prepare RNA Wash Buffer II by adding ethanol according to the instructions Loss of magnetic beads during operation Increase the beads collection time GFC Buffer not diluted wit...

Страница 18: ...16 Notes...

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