Cat. No. IVD3020-20 / IVD3020
Sample-specific RTL or MagZol lysis followed by magnetic RNA binding, on-bead DNase treatment, washing and elution.
Centrifuge 3–5 × 10⁶ suspension cells at 500 × g for 10 min. Remove culture medium completely, loosen the pellet, add 500–600 µL RTL Lysis Buffer and vortex vigorously.
Complete removal of culture medium helps avoid dilution of the chaotropic lysis chemistry.
Add 30 µL MagPure RNA Particles and 500 µL Buffer MCB to a clean 1.5 mL tube, then transfer 500 µL lysate or clarified supernatant from sample preparation.
Prepare Buffer MCB with isopropanol before use.
Mix up and down 15–30 times, incubate at room temperature for 10 min with several intermittent mixes, place on a magnetic rack for 2 min and remove the supernatant.
This step captures RNA on the silica-coated magnetic particles under chaotropic binding conditions.
Add 500 µL Buffer MW1, vortex for 10 s to resuspend the particles, place on the magnetic rack for 1 min and remove the supernatant.
Ensure Buffer MW1 has been prepared with ethanol before use.
Briefly spin to collect liquid from the tube wall, remove all residual liquid carefully and air-dry for 5 min.
This short drying step reduces carryover before on-bead DNase treatment.
Add 300 µL DNase mixture, prepared as 290 µL DNase Buffer plus 10 µL DNase I. Shake gently to resuspend the particles and incubate at room temperature for 10–15 min.
The displayed timeline uses the midpoint of the 10–15 min DNase range.
Add 450 µL Buffer MCB, vortex for 10 s, incubate at room temperature for 5 min, mix up and down 2–3 times, place on the magnetic rack for 1 min and remove the supernatant.
This step restores RNA-binding conditions after DNase treatment.
Add 500 µL Buffer MW1, vortex for 10 s, place on the magnetic rack for 1 min and remove the supernatant.
Complete resuspension improves removal of salts, proteins and DNase reaction components.
Add 500 µL Buffer MW2, vortex for 10 s, place on the magnetic rack for 1 min and remove the supernatant.
MW2 removes residual salts under ethanol-containing wash conditions.
Repeat the MW2 wash once.
Remove the wash buffer completely after magnetic separation.
Briefly spin to collect liquid from the tube wall, remove all residual liquid carefully and air-dry for 10–15 min.
The timeline uses the midpoint of the 10–15 min drying range. Avoid over-drying the particles before elution.
Add 50–100 µL RNase Free Water, vortex to resuspend the particles and incubate at room temperature for 3 min.
Use an elution volume appropriate for the expected RNA concentration and downstream assay.
Place the tube on the magnetic rack for 3 min, then transfer the purified RNA to a new 1.5 mL tube. Store RNA at −20°C or −80°C.
Avoid transferring magnetic particles with the eluate.
Lyse 3–5 × 10⁶ adherent cells directly in the culture dish by adding 600 µL RTL Lysis Buffer and pipetting the lysate several times through a blue pipette tip.
Use additional RTL Lysis Buffer if the lysate is too viscous to aspirate smoothly.
Add 30 µL MagPure RNA Particles and 500 µL Buffer MCB to a clean 1.5 mL tube, then transfer 500 µL lysate or clarified supernatant from sample preparation.
Prepare Buffer MCB with isopropanol before use.
Mix up and down 15–30 times, incubate at room temperature for 10 min with several intermittent mixes, place on a magnetic rack for 2 min and remove the supernatant.
This step captures RNA on the silica-coated magnetic particles under chaotropic binding conditions.
Add 500 µL Buffer MW1, vortex for 10 s to resuspend the particles, place on the magnetic rack for 1 min and remove the supernatant.
Ensure Buffer MW1 has been prepared with ethanol before use.
Briefly spin to collect liquid from the tube wall, remove all residual liquid carefully and air-dry for 5 min.
This short drying step reduces carryover before on-bead DNase treatment.
Add 300 µL DNase mixture, prepared as 290 µL DNase Buffer plus 10 µL DNase I. Shake gently to resuspend the particles and incubate at room temperature for 10–15 min.
The displayed timeline uses the midpoint of the 10–15 min DNase range.
Add 450 µL Buffer MCB, vortex for 10 s, incubate at room temperature for 5 min, mix up and down 2–3 times, place on the magnetic rack for 1 min and remove the supernatant.
This step restores RNA-binding conditions after DNase treatment.
Add 500 µL Buffer MW1, vortex for 10 s, place on the magnetic rack for 1 min and remove the supernatant.
Complete resuspension improves removal of salts, proteins and DNase reaction components.
Add 500 µL Buffer MW2, vortex for 10 s, place on the magnetic rack for 1 min and remove the supernatant.
MW2 removes residual salts under ethanol-containing wash conditions.
Repeat the MW2 wash once.
Remove the wash buffer completely after magnetic separation.
Briefly spin to collect liquid from the tube wall, remove all residual liquid carefully and air-dry for 10–15 min.
The timeline uses the midpoint of the 10–15 min drying range. Avoid over-drying the particles before elution.
Add 50–100 µL RNase Free Water, vortex to resuspend the particles and incubate at room temperature for 3 min.
Use an elution volume appropriate for the expected RNA concentration and downstream assay.
Place the tube on the magnetic rack for 3 min, then transfer the purified RNA to a new 1.5 mL tube. Store RNA at −20°C or −80°C.
Avoid transferring magnetic particles with the eluate.
Homogenize no more than 20 mg animal tissue in 600 µL RTL Lysis Buffer using a mechanical homogenizer, or pulverize in liquid nitrogen and add RTL Lysis Buffer. Centrifuge the lysate at 14,000 × g for 3 min at room temperature.
The timeline assumes a small, well-prepared tissue input. Fibrous or incompletely disrupted tissue may require longer homogenization.
Add 30 µL MagPure RNA Particles and 500 µL Buffer MCB to a clean 1.5 mL tube, then transfer 500 µL lysate or clarified supernatant from sample preparation.
Prepare Buffer MCB with isopropanol before use.
Mix up and down 15–30 times, incubate at room temperature for 10 min with several intermittent mixes, place on a magnetic rack for 2 min and remove the supernatant.
This step captures RNA on the silica-coated magnetic particles under chaotropic binding conditions.
Add 500 µL Buffer MW1, vortex for 10 s to resuspend the particles, place on the magnetic rack for 1 min and remove the supernatant.
Ensure Buffer MW1 has been prepared with ethanol before use.
Briefly spin to collect liquid from the tube wall, remove all residual liquid carefully and air-dry for 5 min.
This short drying step reduces carryover before on-bead DNase treatment.
Add 300 µL DNase mixture, prepared as 290 µL DNase Buffer plus 10 µL DNase I. Shake gently to resuspend the particles and incubate at room temperature for 10–15 min.
The displayed timeline uses the midpoint of the 10–15 min DNase range.
Add 450 µL Buffer MCB, vortex for 10 s, incubate at room temperature for 5 min, mix up and down 2–3 times, place on the magnetic rack for 1 min and remove the supernatant.
This step restores RNA-binding conditions after DNase treatment.
Add 500 µL Buffer MW1, vortex for 10 s, place on the magnetic rack for 1 min and remove the supernatant.
Complete resuspension improves removal of salts, proteins and DNase reaction components.
Add 500 µL Buffer MW2, vortex for 10 s, place on the magnetic rack for 1 min and remove the supernatant.
MW2 removes residual salts under ethanol-containing wash conditions.
Repeat the MW2 wash once.
Remove the wash buffer completely after magnetic separation.
Briefly spin to collect liquid from the tube wall, remove all residual liquid carefully and air-dry for 10–15 min.
The timeline uses the midpoint of the 10–15 min drying range. Avoid over-drying the particles before elution.
Add 50–100 µL RNase Free Water, vortex to resuspend the particles and incubate at room temperature for 3 min.
Use an elution volume appropriate for the expected RNA concentration and downstream assay.
Place the tube on the magnetic rack for 3 min, then transfer the purified RNA to a new 1.5 mL tube. Store RNA at −20°C or −80°C.
Avoid transferring magnetic particles with the eluate.
Disrupt up to 100 mg plant tissue under liquid nitrogen, transfer the powder to a 1.5 mL tube, add 600 µL RTL Lysis Buffer and vortex vigorously. Centrifuge at 14,000 × g for 3 min at room temperature.
Plant samples may vary strongly in polysaccharide and polyphenol content; use a clarified lysate for downstream magnetic binding.
Add 30 µL MagPure RNA Particles and 500 µL Buffer MCB to a clean 1.5 mL tube, then transfer 500 µL lysate or clarified supernatant from sample preparation.
Prepare Buffer MCB with isopropanol before use.
Mix up and down 15–30 times, incubate at room temperature for 10 min with several intermittent mixes, place on a magnetic rack for 2 min and remove the supernatant.
This step captures RNA on the silica-coated magnetic particles under chaotropic binding conditions.
Add 500 µL Buffer MW1, vortex for 10 s to resuspend the particles, place on the magnetic rack for 1 min and remove the supernatant.
Ensure Buffer MW1 has been prepared with ethanol before use.
Briefly spin to collect liquid from the tube wall, remove all residual liquid carefully and air-dry for 5 min.
This short drying step reduces carryover before on-bead DNase treatment.
Add 300 µL DNase mixture, prepared as 290 µL DNase Buffer plus 10 µL DNase I. Shake gently to resuspend the particles and incubate at room temperature for 10–15 min.
The displayed timeline uses the midpoint of the 10–15 min DNase range.
Add 450 µL Buffer MCB, vortex for 10 s, incubate at room temperature for 5 min, mix up and down 2–3 times, place on the magnetic rack for 1 min and remove the supernatant.
This step restores RNA-binding conditions after DNase treatment.
Add 500 µL Buffer MW1, vortex for 10 s, place on the magnetic rack for 1 min and remove the supernatant.
Complete resuspension improves removal of salts, proteins and DNase reaction components.
Add 500 µL Buffer MW2, vortex for 10 s, place on the magnetic rack for 1 min and remove the supernatant.
MW2 removes residual salts under ethanol-containing wash conditions.
Repeat the MW2 wash once.
Remove the wash buffer completely after magnetic separation.
Briefly spin to collect liquid from the tube wall, remove all residual liquid carefully and air-dry for 10–15 min.
The timeline uses the midpoint of the 10–15 min drying range. Avoid over-drying the particles before elution.
Add 50–100 µL RNase Free Water, vortex to resuspend the particles and incubate at room temperature for 3 min.
Use an elution volume appropriate for the expected RNA concentration and downstream assay.
Place the tube on the magnetic rack for 3 min, then transfer the purified RNA to a new 1.5 mL tube. Store RNA at −20°C or −80°C.
Avoid transferring magnetic particles with the eluate.
Lyse the sample with 600 µL Trizol/MagZol Reagent according to the sample route, then centrifuge at 12,000 × g for 10 min at 2–8°C and use the clarified lysate or supernatant for purification.
This route keeps the MagZol front-end but avoids a separate chloroform phase-separation step.
Add 30 µL MagPure RNA Particles and 500 µL Buffer MCB to a clean 1.5 mL tube, then transfer 500 µL lysate or clarified supernatant from sample preparation.
Prepare Buffer MCB with isopropanol before use.
Mix up and down 15–30 times, incubate at room temperature for 10 min with several intermittent mixes, place on a magnetic rack for 2 min and remove the supernatant.
This step captures RNA on the silica-coated magnetic particles under chaotropic binding conditions.
Add 500 µL Buffer MW1, vortex for 10 s to resuspend the particles, place on the magnetic rack for 1 min and remove the supernatant.
Ensure Buffer MW1 has been prepared with ethanol before use.
Briefly spin to collect liquid from the tube wall, remove all residual liquid carefully and air-dry for 5 min.
This short drying step reduces carryover before on-bead DNase treatment.
Add 300 µL DNase mixture, prepared as 290 µL DNase Buffer plus 10 µL DNase I. Shake gently to resuspend the particles and incubate at room temperature for 10–15 min.
The displayed timeline uses the midpoint of the 10–15 min DNase range.
Add 450 µL Buffer MCB, vortex for 10 s, incubate at room temperature for 5 min, mix up and down 2–3 times, place on the magnetic rack for 1 min and remove the supernatant.
This step restores RNA-binding conditions after DNase treatment.
Add 500 µL Buffer MW1, vortex for 10 s, place on the magnetic rack for 1 min and remove the supernatant.
Complete resuspension improves removal of salts, proteins and DNase reaction components.
Add 500 µL Buffer MW2, vortex for 10 s, place on the magnetic rack for 1 min and remove the supernatant.
MW2 removes residual salts under ethanol-containing wash conditions.
Repeat the MW2 wash once.
Remove the wash buffer completely after magnetic separation.
Briefly spin to collect liquid from the tube wall, remove all residual liquid carefully and air-dry for 10–15 min.
The timeline uses the midpoint of the 10–15 min drying range. Avoid over-drying the particles before elution.
Add 50–100 µL RNase Free Water, vortex to resuspend the particles and incubate at room temperature for 3 min.
Use an elution volume appropriate for the expected RNA concentration and downstream assay.
Place the tube on the magnetic rack for 3 min, then transfer the purified RNA to a new 1.5 mL tube. Store RNA at −20°C or −80°C.
Avoid transferring magnetic particles with the eluate.
Lyse the sample with 1 mL Trizol/MagZol Reagent, add 200 µL chloroform to the lysate, then centrifuge at 12,000 × g for 15 min at 2–8°C before transferring the aqueous phase or clarified supernatant.
Avoid disturbing the interphase when collecting the RNA-containing phase.
Add 30 µL MagPure RNA Particles and 500 µL Buffer MCB to a clean 1.5 mL tube, then transfer 500 µL lysate or clarified supernatant from sample preparation.
Prepare Buffer MCB with isopropanol before use.
Mix up and down 15–30 times, incubate at room temperature for 10 min with several intermittent mixes, place on a magnetic rack for 2 min and remove the supernatant.
This step captures RNA on the silica-coated magnetic particles under chaotropic binding conditions.
Add 500 µL Buffer MW1, vortex for 10 s to resuspend the particles, place on the magnetic rack for 1 min and remove the supernatant.
Ensure Buffer MW1 has been prepared with ethanol before use.
Briefly spin to collect liquid from the tube wall, remove all residual liquid carefully and air-dry for 5 min.
This short drying step reduces carryover before on-bead DNase treatment.
Add 300 µL DNase mixture, prepared as 290 µL DNase Buffer plus 10 µL DNase I. Shake gently to resuspend the particles and incubate at room temperature for 10–15 min.
The displayed timeline uses the midpoint of the 10–15 min DNase range.
Add 450 µL Buffer MCB, vortex for 10 s, incubate at room temperature for 5 min, mix up and down 2–3 times, place on the magnetic rack for 1 min and remove the supernatant.
This step restores RNA-binding conditions after DNase treatment.
Add 500 µL Buffer MW1, vortex for 10 s, place on the magnetic rack for 1 min and remove the supernatant.
Complete resuspension improves removal of salts, proteins and DNase reaction components.
Add 500 µL Buffer MW2, vortex for 10 s, place on the magnetic rack for 1 min and remove the supernatant.
MW2 removes residual salts under ethanol-containing wash conditions.
Repeat the MW2 wash once.
Remove the wash buffer completely after magnetic separation.
Briefly spin to collect liquid from the tube wall, remove all residual liquid carefully and air-dry for 10–15 min.
The timeline uses the midpoint of the 10–15 min drying range. Avoid over-drying the particles before elution.
Add 50–100 µL RNase Free Water, vortex to resuspend the particles and incubate at room temperature for 3 min.
Use an elution volume appropriate for the expected RNA concentration and downstream assay.
Place the tube on the magnetic rack for 3 min, then transfer the purified RNA to a new 1.5 mL tube. Store RNA at −20°C or −80°C.
Avoid transferring magnetic particles with the eluate.
Centrifuge up to 10 mL urine at 1,600 × g for 10 min to collect cast-off cells. Remove the supernatant, add 500–600 µL RTL Lysis Buffer and vortex vigorously to resuspend the pellet.
The useful RNA fraction depends on the recovered cell pellet; low-cell urine samples may give limited RNA input.
Add 30 µL MagPure RNA Particles and 500 µL Buffer MCB to a clean 1.5 mL tube, then transfer 500 µL lysate or clarified supernatant from sample preparation.
Prepare Buffer MCB with isopropanol before use.
Mix up and down 15–30 times, incubate at room temperature for 10 min with several intermittent mixes, place on a magnetic rack for 2 min and remove the supernatant.
This step captures RNA on the silica-coated magnetic particles under chaotropic binding conditions.
Add 500 µL Buffer MW1, vortex for 10 s to resuspend the particles, place on the magnetic rack for 1 min and remove the supernatant.
Ensure Buffer MW1 has been prepared with ethanol before use.
Briefly spin to collect liquid from the tube wall, remove all residual liquid carefully and air-dry for 5 min.
This short drying step reduces carryover before on-bead DNase treatment.
Add 300 µL DNase mixture, prepared as 290 µL DNase Buffer plus 10 µL DNase I. Shake gently to resuspend the particles and incubate at room temperature for 10–15 min.
The displayed timeline uses the midpoint of the 10–15 min DNase range.
Add 450 µL Buffer MCB, vortex for 10 s, incubate at room temperature for 5 min, mix up and down 2–3 times, place on the magnetic rack for 1 min and remove the supernatant.
This step restores RNA-binding conditions after DNase treatment.
Add 500 µL Buffer MW1, vortex for 10 s, place on the magnetic rack for 1 min and remove the supernatant.
Complete resuspension improves removal of salts, proteins and DNase reaction components.
Add 500 µL Buffer MW2, vortex for 10 s, place on the magnetic rack for 1 min and remove the supernatant.
MW2 removes residual salts under ethanol-containing wash conditions.
Repeat the MW2 wash once.
Remove the wash buffer completely after magnetic separation.
Briefly spin to collect liquid from the tube wall, remove all residual liquid carefully and air-dry for 10–15 min.
The timeline uses the midpoint of the 10–15 min drying range. Avoid over-drying the particles before elution.
Add 50–100 µL RNase Free Water, vortex to resuspend the particles and incubate at room temperature for 3 min.
Use an elution volume appropriate for the expected RNA concentration and downstream assay.
Place the tube on the magnetic rack for 3 min, then transfer the purified RNA to a new 1.5 mL tube. Store RNA at −20°C or −80°C.
Avoid transferring magnetic particles with the eluate.
The official protocol also provides 32/48-channel and 96-channel extractor routes. In the automated setup, 400–450 µL lysate or clarified supernatant is loaded into the plate with Buffer MCB, MagPure RNA Particles are placed in the MW1 wash position, and DNase Buffer plus DNase I is loaded in the DNase position. The program pauses at approximately 25 min so that 450 µL Buffer MCB can be added to the DNase well or plate before the run continues. This workflow note uses the manual magnetic-bead route for the standard cumulative timeline.
This workflow separates sample-specific pretreatment from the shared downstream purification path. It is intended as a practical companion to the product manual rather than a replacement for the official protocol. Where multiple protocol routes are available, this note focuses on the route that best represents the key workflow logic of this product.
Protocol times stated in the product manual are retained where applicable. Steps without explicit timing are estimated for an experienced operator, including pipetting, tube transfer, centrifuge or magnetic-rack handling, supernatant / filtrate removal and tube, plate or column repositioning. For short protocol ranges, the timeline uses the midpoint. For long or optional protocol ranges, the displayed standard timeline uses the shortest reasonable path, while the note and total-time range indicate where extended handling may apply. Cumulative time is calculated separately within each sample-specific tab, running continuously from that tab’s first preparation step to final elution.
IVD3020 is a magnetic-particle total RNA workflow with integrated DNase I treatment. RNA is first captured from RTL lysate or compatible MagZol-derived lysate under Buffer MCB binding conditions. DNA is then reduced by on-bead DNase digestion before the RNA is rebound, washed with MW1 and MW2, dried and eluted in RNase Free Water.
Use the correct front-end route for the sample matrix and control input amount carefully, especially for tissue, plant powder, viscous cell pellets and urine cell pellets. Clarified lysate or supernatant should be used for binding. MW1 and MW2 must be prepared with ethanol, Buffer MCB must be prepared with isopropanol, and optional 2-mercaptoethanol or DTT can be added to RTL Lysis Buffer when stronger RNase control is required. During drying, remove residual wash buffer completely but avoid excessive over-drying before elution.