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32. Transfer the cleared supernatant containing purified RNA into a new RNase-free

microplate.

33. Store eluted RNA at -70°C.

Note:

Any combination of the following steps can be used to help increase RNA yield.

•

Heat the DEPC Water to 70°C before adding to the beads.

•

Increase the incubation time to 5 minutes.

•

Increase the elution volume.

•

Repeat the elution step with fresh DEPC Water (this may increase the yield, but
decrease the concentration).

M2837 Mag-Bind® Blood RNA 96 Protocol

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Содержание Mag-Bind M2837-00

Страница 1: ...Mag Bind Blood RNA 96 Kit 50 L Blood M2837 00 1 x 96 preps M2837 01 4 x 96 preps 200 L Blood M2839 00 1 x 96 preps M2839 01 4 x 96 preps May 2016...

Страница 2: ...nts Storage and Stability 5 Preparing Reagents 6 50 L Mag Bind Blood RNA 96 Protocol M2837 7 200 L Mag Bind Blood RNA 96 Protocol M2839 12 Troubleshooting Guide 17 Ordering 18 Manual Revision May 2016...

Страница 3: ...omplex is separated from lysates using a magnet Proteins and cellular debris are efficiently washed away by a washing step Next DNA is removed with a Mag Bind DNase I treatment RNA is rebound and clea...

Страница 4: ...gloves to minimize RNase contamination Use only clean RNase free disposable plastic pipette tips and plastic ware for the supplied reagents Equilibrate samples and reagents to room temperature before...

Страница 5: ...r Under these conditions RNA is stable for more than a year Integrity of RNA It is highly recommended that RNA quality be determined prior to beginning all downstream applications The quality of RNA c...

Страница 6: ...on 2 4 mL 9 mL RXT Wash Buffer 55 mL 220 mL RNA Wash Buffer II 50 mL 2 x 70 mL DEPC Water 15 mL 60 mL Mag Bind DNase I 225 L 900 L DNase l Digestion Buffer 12 mL 48 mL User Manual P P Storage and Stab...

Страница 7: ...37 00 100 mL M2837 01 400 mL Kit 100 Ethanol to be Added M2839 00 200 mL M2839 01 280 mL per bottle 2 Dilute RXT Wash Buffer with 100 ethanol as follows and store at room temperature Kit 100 Ethanol t...

Страница 8: ...A Wash Buffer II and RXT Wash Buffer according to the Preparing Reagents section on Page 6 1 Add 65 L RBL Buffer and 65 L isopropanol to each well of a 500 L 96 well microplate 2 Add 50 L blood sample...

Страница 9: ...10 Aspirate and discard the cleared supernatant Do not disturb the Mag Bind Particles CNR 11 Remove the plate containing the Mag Bind Particles CNR from the magnetic separation device 12 Repeat Steps...

Страница 10: ...ing error Important Notes Mag Bind DNase I is very sensitive and prone to physical denaturation Do not vortex the Mag Bind DNase I mixture Mix gently by shaking the plate Freshly prepare Mag Bind DNas...

Страница 11: ...etic separation device to magnetize the Mag Bind Particles CNR Let sit at room temperature until the Mag Bind Particles CNR are completely cleared from solution 27 Aspirate and discard the cleared sup...

Страница 12: ...ination of the following steps can be used to help increase RNA yield Heat the DEPC Water to 70 C before adding to the beads Increase the incubation time to 5 minutes Increase the elution volume Repea...

Страница 13: ...Cat EZ9604 01 Multichannel pipette Nuclease free pipette tips 100 ethanol Isopropanol Sealing film Before Starting Prepare RNA Wash Buffer II and RXT Wash Buffer according to the Preparing Reagents s...

Страница 14: ...e to magnetize the Mag Bind Particles CNR Let sit at room temperature until the Mag Bind Particles CNR are completely cleared from solution 10 Aspirate and discard the cleared supernatant Do not distu...

Страница 15: ...8 L 22 L 1100 L 96 10 35 mL 211 L 10 56 mL Volumes are calculated 10 extra to offset pipetting error Important Notes Mag Bind DNase I is very sensitive and prone to physical denaturation Do not vortex...

Страница 16: ...5 Add 450 L RNA Wash Buffer II to each well Pipet up and down 20 times or shake for 2 minutes to mix thoroughly 26 Place the plate on a magnetic separation device to magnetize the Mag Bind Particles C...

Страница 17: ...taining purified RNA into a new RNase free microplate 33 Store eluted RNA at 70 C Note Any combination of the following steps can be used to help increase RNA yield Heat the DEPC Water to 70 C before...

Страница 18: ...sample is clear of blood clots before adding magnetic beads Problem Cause Solution No RNA eluted RNA Wash Buffer II was not diluted with 100 ethanol Prepare RNA Wash Buffer II by adding ethanol accor...

Страница 19: ...MSD 02 96 well Square well Plate 2 2 mL 5 pk EZ9602 01 96 well Round well Plate 1 2 mL 10 pk SSI 1780 00 96 well Microplate 500 L EZ9604 01 DEPC Water 30 mL PR032 RNA Wash Buffer II 20 mL PDR046 HiBin...

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