Omega Bio-Tek Mag-Bind RXNPure Plus User manual

Mag-Bind® RXNPure Plus
M1386-01 50 mL
Manual Date: February 2023
Revision Number: v4.0

1
Introduction ....................................................................................2
Illustrated Protocol........................................................................3
Kit Contents/Preparations/Storage andStability................4
Mag-Bind® RXNPure Plus 96-well Plate Protocol................5
Mag-Bind® RXNPurePlus 384-wellPlateProtocol................8
Troubleshooting Guide.............................................................11
Manual Date: February 2023
Revision Number: v4.0
Mag-Bind® RXNPure Plus
Table of Contents

2
Introduction
Omega Bio-tek’s Mag-Bind® RXNPure Plus Kit allows rapid and reliable isolation of PCR
products with high recovery rates. The system combines Omega Bio-tek’s proprietary
chemistries with the reversible nucleic acid-binding properties of magnetic beads that
selectively binds PCR amplicons 100 bp and larger and eliminates excess nucleotides,
primers, and small, non-targeted amplication products, such as primer dimers. This kit
is designed for both manual and fully automated purication of PCR samples. Puried
PCR fragments can be used for microarrays, automated uorescent DNA sequencing,
restriction enzyme digestion, and other applications.
The Mag-Bind® RXNPure Plus magnetic particles technology provides a better solution
for nucleic acid purication compared to centrifugation and vacuum-based technologies.
The product can be easily scaled up while providing very user-friendly handling
procedures. If using Mag-Bind® RXNPure Plus for the rst time, please read this booklet
to become familiar with the procedures. PCR products are rst mixed with Mag-Bind®
RXNPure Plus. PCR products then selectively bind to the Mag-Bind® RXNPure Plus
particles. With two rapid wash steps, trace contaminants such as nucleotides, primers and
small, non-targeted amplication products are removed. Pure DNA is eluted in Elution
Buer or water. Puried DNA can be directly used in downstream applications without the
need for further purication.
Important: If automating this procedure on a liquid handler or a magnetic processor,
please contact your Omega Bio-tek representative for instrument-specic instructions.
New in this Edition:
February 2023
• M1386-00 (5 mL) and M1386-02 (500 mL) have been discontinued and are no longer
available to purchase.
March 2017
• General Revision

3
Illustrated Protocol
Measure the PCR Reaction
Add Mag-Bind®RXNPure Plus and Mix
Magnetize and Remove Supernatant
Wash Twice with 70% Ethanol
Dry
Elute DNA

4
Kit Contents
Storage and Stability
Mag-Bind® RXNPure Plus is guaranteed for at least 12 months from the date of purchase
when stored at 2-8°C.
Product Number M1386-01
Mag-Bind® RXNPure Plus 50 mL
User Manual P
Preparations
PCR Reaction Volume
96 well format
M1386-01
50 mL
10 μL 2,777 preps
25 μL 1,111 preps
50 μL 555 preps
100 μL 277 preps
PCR Reaction Volume
384 well format
M1386-01
50 mL
5 μL 5,555 preps
10 μL 2,777 preps
14 μL 1,984 preps

5
Mag-Bind® RXNPure Plus
96-well Plate Protocol
Important: If automating this procedure on a liquid handler or a magnetic processor,
please contact your Omega Bio-tek representative for instrument-specic instructions.
Materials and Equipment to be Supplied by User:
• Vortexer
• Magnetic Separation Device (Recommended Cat# AlpAqua A001322)
• Multichannel pipettor
• 96-well PCR plate containing PCR samples (up to 50 μL/well)
• 96-well microplate
• Multichannel Disposable Reservoirs
• Sealing lm
• 70% ethanol
• Elution Buer (Cat# PDR048 or 10 mM Tris, pH 8.5)
• Optional: Oven capable of 37°C
1. Read the manufacturer’s instruction manual for the magnetic separation device, if
provided.
2. Place the 96-well PCR plate containing PCR samples on the bench and measure the
volume of the PCR reaction. Determine the volume of Mag-Bind® RXNPure Plus that
will be added to the reaction. If the reaction volume will exceed 200 µL transfer to a
microtiter plate for processing.
Note: PCR reactions >20 µL will need to be transferred to a processing plate.
If processing in a PCR plate, a magnet separation device compatible with PCR plates
must be used (Recommend Cat# V&P Scientic VP 771H).
3. Shake or vortex the Mag-Bind® RXNPure Plus to resuspend any particles that may
have settled. Allow Mag-Bind® RXNPure Plus to come to room temperature before
use.

6
4. Add 1.8 volumes Mag-Bind® RXNPure Plus.
PCR Reaction Volume (μL) Mag-Bind®RXNPure Plus (μL)
10 18
25 45
50 90
100 180
5. Pipet up and down 5-10 times or vortex for 30 seconds.
6. Let sit for 5 minutes at room temperature.
7. Place the plate on a magnetic separation device to magnetize the Mag-Bind®
RXNPure Plus. Let sit at room temperature until the Mag-Bind® RXNPure Plus is
completely cleared from solution.
8. Aspirate and discard the cleared supernatant. Do not disturb the Mag-Bind® RXNPure
Plus. Leave the plate on the magnet.
9. Add 200 μL 70% ethanol.
10. Let sit at room temperature for 1 minute. It is not necessary to resuspend the Mag-
Bind® RXNPure Plus.
11. Aspirate and discard the cleared supernatant. Do not disturb the Mag-Bind® RXNPure
Plus.
12. Repeat Steps 9-11 for a second 70% ethanol wash step.
Mag-Bind® RXNPure Plus

7
13. Leave the plate on the magnetic separation device for 10-15 minutes to air dry the
Mag-Bind® RXNPure Plus. Remove any residual liquid with a pipettor.
Note: It is important to dry the Mag-Bind® RXNPure Plus before elution. Residual
ethanol may interfere with downstream applications.
Optional: Incubating the plate at 37°C can speed up the evaporation.
14. Remove the plate from magnetic separation device.
15. Add 30-40 µL Elution Buer (not provided).
16. Pipet up and down 20 times or vortex for 30 seconds.
17. Let sit for 2-3 minutes at room temperature.
18. Place the plate on a magnetic separation device to magnetize the Mag-Bind®
RXNPure Plus. Let sit at room temperature until the Mag-Bind® RXNPure Plus is
completely cleared from solution.
19. Transfer the cleared supernatant containing puried DNA to a new 96-well
microplate and seal with non-permeable sealing lm.
20. Store the plate at 2-8°C if storage is only for a few days. For long-term storage,
samples should be kept at -20°C.
Mag-Bind® RXNPure Plus

8
Mag-Bind® RXNPure Plus
384-well Plate Protocol
Important: If automating this procedure on a liquid handler or a magnetic processor,
please contact your Omega Bio-tek representative for instrument-specic instructions.
Materials and Equipment to be Supplied by User:
• Vortexer
• Magnetic separation device for 384-well PCR plates
• Multichannel pipettor
• 384-well PCR plate containing PCR samples (up to 100 μL/well)
• Skirted 384-well PCR plate
• 384-well microplate
• Multichannel Disposable Reservoirs
• Sealing lm
• 70% ethanol
• Elution Buer (Cat# PDR048 or 10 mM Tris, pH 8.5)
• Optional: Oven capable of 37°C
1. Read the manufacturer’s instruction manual for the magnetic separation device, if
provided.
2. Place the 384-well PCR plate on the bench and measure the volume of the PCR
reaction. Transfer the sample to a skirted 384-well PCR plate.
3. Shake or vortex the Mag-Bind® RXNPure Plus to resuspend any particles that may
have settled. Allow Mag-Bind® RXNPure Plus to come to room temperature before
use.
4. Add 1.8 volumes Mag-Bind® RXNPure Plus.
PCR Reaction Volume (μL) Mag-Bind®RXNPure Plus (μL)
5 9
10 18
14 25

9
5. Pipet up and down 5-10 times or vortex for 30 seconds.
6. Let sit for 1 minute at room temperature.
7. Place the plate on a magnetic separation device to magnetize the Mag-Bind®
RXNPure Plus. Let sit at room temperature until the Mag-Bind® RXNPure Plus is
completely cleared from solution.
8. Aspirate and discard the cleared supernatant. Do not disturb the Mag-Bind® RXNPure
Plus. Leave the plate on the magnet.
9. Add 30 μL 70% ethanol.
10. Let sit at room temperature for 1 minute. It is not necessary to resuspend the Mag-
Bind® RXNPure Plus.
11. Aspirate and discard the cleared supernatant. Do not disturb the Mag-Bind® RXNPure
Plus.
12. Repeat Steps 9-11 for a second 70% ethanol wash step.
13. Leave the plate on the magnetic separation device for 10-15 minutes to air dry the
Mag-Bind® RXNPure Plus. Remove any residual liquid with a pipettor.
Note: It is important to dry the Mag-Bind® RXNPure Plus before elution. Residual
ethanol may interfere with downstream applications.
Optional: Incubating the plate at 37°C can speed up the evaporation.
14. Remove the plate from magnetic separation device.
15. Add 30 µL Elution Buer (not provided).
Mag-Bind® RXNPure Plus

10
16. Pipet up and down 20 times or vortex for 30 seconds.
17. Let sit for 2-3 minutes at room temperature.
18. Place the plate on a magnetic separation device to magnetize the Mag-Bind®
RXNPure Plus. Let sit at room temperature until the Mag-Bind® RXNPure Plus is
completely cleared from solution.
19. Transfer the cleared supernatant containing puried DNA to a new 384-well
microplate and seal with non-permeable sealing lm.
20. Store the plate at 2-8°C if storage is only for a few days. For long-term storage,
samples should be kept at -20°C.
Mag-Bind® RXNPure Plus

11
Troubleshooting Guide
Please use this guide to troubleshoot any problems that may arise. For further assistance,
please contact the technical support sta, toll free, at 800-832-8896.
Possible Problems and Suggestions
Problem Cause Solution
Low yield
Low PCR product yield Increase the number amplication cycles
for PCR.
Smaller PCR product size Small PCR fragments normally give lower
yield.
Ethanol residue During the drying step, remove any
liquid from bottom of the well.
Particle loss during the
procedure
Increase magnetization time.
Aspirate slowly.
DNA remains bound to
beads Increase elution volume.
Incomplete resuspension
of the beads during
elution
Vortex or pipet up and down to fully
resuspend the beads.
Problem Cause Solution
Primer
carryover
Insucient wash of the
particles
Wash the beads one more time with 70%
ethanol.
Problem Solution
Non-specic
amplication
products were
not removed
The size of the non-
specic amplication
products are larger than
100 bp
Non-specic amplication products
larger than 100 bp are not eciently
removed from PCR products.
Problem Cause Solution
Problems in
downstream
applications
Salt carryover 70% ethanol must be stored at room
temperature.
Ethanol carryover Ensure the beads are completely dried
before elution.

12
Notes: