Cat. No. R662300 / R662301 / R662302 / R662303
Sample-specific MagZol lysis followed by BCP phase separation and magnetic-particle RNA purification.
Homogenize 10–50 mg animal tissue or 10–100 mg blood vessel in 1 mL MagZol Reagent using a pestle or homogenizer.
For fibrous vascular samples, continue homogenization until the lysate is evenly dispersed before BCP addition.
Add 100 µL Buffer BCP per 1 mL MagZol lysate. Shake vigorously by hand for 15 s and incubate at room temperature for 3 min.
Do not replace vigorous hand shaking with vortexing; the manual warns that vortex mixing may increase DNA contamination.
Centrifuge at 12,000 × g for 15 min at 4°C and recover the RNA-containing upper aqueous phase/supernatant for purification.
Use the clarified upper phase; avoid disturbing the interphase.
In a 1.5 mL tube, combine 500 µL isopropanol, 30 µL MagPure Particles N and 500 µL phase-separated supernatant. Invert 15–20 times, incubate at room temperature for 10 min with intermittent mixing, place on a magnetic rack for 2 min, then remove supernatant.
This is the main RNA capture step after MagZol/BCP phase separation.
Add 500 µL Buffer MW1, vortex for 15 s, place on the magnetic rack for 1 min and remove supernatant.
Ensure Buffer MW1 has been prepared with ethanol before use.
Add 500 µL Buffer MW2, vortex for 15 s, place on the magnetic rack for 1 min and remove supernatant.
This wash removes residual salts and organic contaminants.
Repeat the MW2 wash once.
Remove the supernatant completely after magnetic separation.
Briefly centrifuge to collect liquid from the tube wall, remove all residual reagent, then dry at room temperature for 10 min.
Avoid over-drying the particles; residual ethanol can inhibit downstream RT-PCR.
Add 30–100 µL RNase Free Water, vortex to disperse the beads, incubate at room temperature for 5 min, place on magnetic rack for 3 min, then transfer purified RNA to a clean tube.
Store RNA at −20°C or −80°C after elution.
Grind plant material or bone/teeth sample under liquid nitrogen, transfer the recommended powder amount, then immediately add 1 mL MagZol Reagent and vortex thoroughly.
Plant input is typically 30–100 mg powder; bone/teeth powder is typically 100–200 mg.
Add 100 µL Buffer BCP per 1 mL MagZol lysate. Shake vigorously by hand for 15 s and incubate at room temperature for 3 min.
Do not replace vigorous hand shaking with vortexing; the manual warns that vortex mixing may increase DNA contamination.
Centrifuge at 12,000 × g for 15 min at 4°C and recover the RNA-containing upper aqueous phase/supernatant for purification.
Use the clarified upper phase; avoid disturbing the interphase.
In a 1.5 mL tube, combine 500 µL isopropanol, 30 µL MagPure Particles N and 500 µL phase-separated supernatant. Invert 15–20 times, incubate at room temperature for 10 min with intermittent mixing, place on a magnetic rack for 2 min, then remove supernatant.
This is the main RNA capture step after MagZol/BCP phase separation.
Add 500 µL Buffer MW1, vortex for 15 s, place on the magnetic rack for 1 min and remove supernatant.
Ensure Buffer MW1 has been prepared with ethanol before use.
Add 500 µL Buffer MW2, vortex for 15 s, place on the magnetic rack for 1 min and remove supernatant.
This wash removes residual salts and organic contaminants.
Repeat the MW2 wash once.
Remove the supernatant completely after magnetic separation.
Briefly centrifuge to collect liquid from the tube wall, remove all residual reagent, then dry at room temperature for 10 min.
Avoid over-drying the particles; residual ethanol can inhibit downstream RT-PCR.
Add 30–100 µL RNase Free Water, vortex to disperse the beads, incubate at room temperature for 5 min, place on magnetic rack for 3 min, then transfer purified RNA to a clean tube.
Store RNA at −20°C or −80°C after elution.
Remove culture medium completely. Add 1 mL MagZol Reagent per 10 cm² culture area and pipette 3–5 times to fully lyse the cells.
Avoid leaving residual culture medium, which dilutes the MagZol lysis chemistry.
Add 100 µL Buffer BCP per 1 mL MagZol lysate. Shake vigorously by hand for 15 s and incubate at room temperature for 3 min.
Do not replace vigorous hand shaking with vortexing; the manual warns that vortex mixing may increase DNA contamination.
Centrifuge at 12,000 × g for 15 min at 4°C and recover the RNA-containing upper aqueous phase/supernatant for purification.
Use the clarified upper phase; avoid disturbing the interphase.
In a 1.5 mL tube, combine 500 µL isopropanol, 30 µL MagPure Particles N and 500 µL phase-separated supernatant. Invert 15–20 times, incubate at room temperature for 10 min with intermittent mixing, place on a magnetic rack for 2 min, then remove supernatant.
This is the main RNA capture step after MagZol/BCP phase separation.
Add 500 µL Buffer MW1, vortex for 15 s, place on the magnetic rack for 1 min and remove supernatant.
Ensure Buffer MW1 has been prepared with ethanol before use.
Add 500 µL Buffer MW2, vortex for 15 s, place on the magnetic rack for 1 min and remove supernatant.
This wash removes residual salts and organic contaminants.
Repeat the MW2 wash once.
Remove the supernatant completely after magnetic separation.
Briefly centrifuge to collect liquid from the tube wall, remove all residual reagent, then dry at room temperature for 10 min.
Avoid over-drying the particles; residual ethanol can inhibit downstream RT-PCR.
Add 30–100 µL RNase Free Water, vortex to disperse the beads, incubate at room temperature for 5 min, place on magnetic rack for 3 min, then transfer purified RNA to a clean tube.
Store RNA at −20°C or −80°C after elution.
Collect fewer than 5 × 10⁶ cells by centrifugation at 500 × g, remove medium, loosen the pellet, add 1 mL MagZol Reagent and pipette 3–5 times.
Fully disperse the pellet before phase separation; viscous lysates usually indicate excessive input or incomplete mixing.
Add 100 µL Buffer BCP per 1 mL MagZol lysate. Shake vigorously by hand for 15 s and incubate at room temperature for 3 min.
Do not replace vigorous hand shaking with vortexing; the manual warns that vortex mixing may increase DNA contamination.
Centrifuge at 12,000 × g for 15 min at 4°C and recover the RNA-containing upper aqueous phase/supernatant for purification.
Use the clarified upper phase; avoid disturbing the interphase.
In a 1.5 mL tube, combine 500 µL isopropanol, 30 µL MagPure Particles N and 500 µL phase-separated supernatant. Invert 15–20 times, incubate at room temperature for 10 min with intermittent mixing, place on a magnetic rack for 2 min, then remove supernatant.
This is the main RNA capture step after MagZol/BCP phase separation.
Add 500 µL Buffer MW1, vortex for 15 s, place on the magnetic rack for 1 min and remove supernatant.
Ensure Buffer MW1 has been prepared with ethanol before use.
Add 500 µL Buffer MW2, vortex for 15 s, place on the magnetic rack for 1 min and remove supernatant.
This wash removes residual salts and organic contaminants.
Repeat the MW2 wash once.
Remove the supernatant completely after magnetic separation.
Briefly centrifuge to collect liquid from the tube wall, remove all residual reagent, then dry at room temperature for 10 min.
Avoid over-drying the particles; residual ethanol can inhibit downstream RT-PCR.
Add 30–100 µL RNase Free Water, vortex to disperse the beads, incubate at room temperature for 5 min, place on magnetic rack for 3 min, then transfer purified RNA to a clean tube.
Store RNA at −20°C or −80°C after elution.
Use about 1 mL whole blood or 0.5 mL bone marrow. Separate lymphocytes using lymphocyte separation medium or red blood cell lysis buffer, leave 50–100 µL residual liquid with the pellet, then add 1 mL MagZol Reagent.
Control the white-cell pellet size, especially for bone marrow; excessive leukocyte precipitation can increase viscosity and reduce extraction quality.
Add 100 µL Buffer BCP per 1 mL MagZol lysate. Shake vigorously by hand for 15 s and incubate at room temperature for 3 min.
Do not replace vigorous hand shaking with vortexing; the manual warns that vortex mixing may increase DNA contamination.
Centrifuge at 12,000 × g for 15 min at 4°C and recover the RNA-containing upper aqueous phase/supernatant for purification.
Use the clarified upper phase; avoid disturbing the interphase.
In a 1.5 mL tube, combine 500 µL isopropanol, 30 µL MagPure Particles N and 500 µL phase-separated supernatant. Invert 15–20 times, incubate at room temperature for 10 min with intermittent mixing, place on a magnetic rack for 2 min, then remove supernatant.
This is the main RNA capture step after MagZol/BCP phase separation.
Add 500 µL Buffer MW1, vortex for 15 s, place on the magnetic rack for 1 min and remove supernatant.
Ensure Buffer MW1 has been prepared with ethanol before use.
Add 500 µL Buffer MW2, vortex for 15 s, place on the magnetic rack for 1 min and remove supernatant.
This wash removes residual salts and organic contaminants.
Repeat the MW2 wash once.
Remove the supernatant completely after magnetic separation.
Briefly centrifuge to collect liquid from the tube wall, remove all residual reagent, then dry at room temperature for 10 min.
Avoid over-drying the particles; residual ethanol can inhibit downstream RT-PCR.
Add 30–100 µL RNase Free Water, vortex to disperse the beads, incubate at room temperature for 5 min, place on magnetic rack for 3 min, then transfer purified RNA to a clean tube.
Store RNA at −20°C or −80°C after elution.
For bacteria, collect about 1 × 10⁸ cells, treat with 100 µL TE/lysozyme for 10 min, then add 1 mL MagZol Reagent and vortex for 1 min. For trace fungi, lyse <10 mg sample in a fungal/bacterial homogenate tube with 1 mL MagZol Reagent by high-speed vortexing for 5–10 min.
Bacterial and fungal samples depend strongly on cell-wall disruption efficiency; insufficient pre-lysis is the most common cause of low RNA recovery.
Add 100 µL Buffer BCP per 1 mL MagZol lysate. Shake vigorously by hand for 15 s and incubate at room temperature for 3 min.
Do not replace vigorous hand shaking with vortexing; the manual warns that vortex mixing may increase DNA contamination.
Centrifuge at 12,000 × g for 15 min at 4°C and recover the RNA-containing upper aqueous phase/supernatant for purification.
Use the clarified upper phase; avoid disturbing the interphase.
In a 1.5 mL tube, combine 500 µL isopropanol, 30 µL MagPure Particles N and 500 µL phase-separated supernatant. Invert 15–20 times, incubate at room temperature for 10 min with intermittent mixing, place on a magnetic rack for 2 min, then remove supernatant.
This is the main RNA capture step after MagZol/BCP phase separation.
Add 500 µL Buffer MW1, vortex for 15 s, place on the magnetic rack for 1 min and remove supernatant.
Ensure Buffer MW1 has been prepared with ethanol before use.
Add 500 µL Buffer MW2, vortex for 15 s, place on the magnetic rack for 1 min and remove supernatant.
This wash removes residual salts and organic contaminants.
Repeat the MW2 wash once.
Remove the supernatant completely after magnetic separation.
Briefly centrifuge to collect liquid from the tube wall, remove all residual reagent, then dry at room temperature for 10 min.
Avoid over-drying the particles; residual ethanol can inhibit downstream RT-PCR.
Add 30–100 µL RNase Free Water, vortex to disperse the beads, incubate at room temperature for 5 min, place on magnetic rack for 3 min, then transfer purified RNA to a clean tube.
Store RNA at −20°C or −80°C after elution.
The official protocol also provides a 32/48-channel extractor setup. After MagZol / BCP phase separation, 400–500 µL supernatant is added to Row 1/7 containing isopropanol and MagPure Particles N, and program #R6623 is run for approximately 30 min. 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 runs continuously from the first step to final elution across all workflow sections.
R6623 uses a MagZol / BCP phase-separation front end followed by magnetic-particle RNA purification. The main workflow logic is sample disruption in MagZol Reagent, BCP-assisted phase separation to remove genomic DNA and protein impurities, recovery of the clarified RNA-containing upper phase, and RNA capture on MagPure Particles N under isopropanol-assisted binding conditions. This note uses the manual magnetic-bead route as the standard cumulative timeline, while the automation route is summarized separately because it starts after the same phase-separation step.
Front-end lysis quality is strongly sample dependent. Fibrous tissue, plant powder, bone, blood vessels, fungi, bacteria and blood-cell or bone-marrow pellets may require careful homogenization, enzymatic pretreatment or bead-tube disruption before the shared phase-separation step. During BCP mixing, vigorous hand shaking is required; slow inversion may reduce extraction efficiency, while vortex mixing at this step may increase DNA contamination. After centrifugation, transfer only the clarified upper phase and avoid disturbing the interphase. Ensure MW1 and MW2 are prepared with ethanol, remove residual wash buffer thoroughly, and avoid excessive bead over-drying before elution.