One-Step RNA Removal Kit uses Ribonuclease A immobilized magnetic beads to efficiently remove RNA from the sample using a single step protocol. Ribonuclease A is an endoribonuclease that originates from the bovine pancreas. RNase A is a single chain polypeptide with a molecular mass of 13.7 kDa. RNase A is an endoribonuclease that unpacifically degrades ribonucleic acid (RNA) into smaller components. The magnetic bead immobilized with RNase A can efficiently remove RNA from biological samples with no nucleases remaining in the solution due to the nuclease stably and covalently conjugated with the magnetic beads.
Applications
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For analytical purposes
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Plasmid and genomic DNA preparation
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For cell cycle analysis by flow cytometry and propidium iodide (PI) staining
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Removal of RNA from recombinant protein preparations
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Ribonuclease protection assays. Used in conjunction with RNase T1
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Mapping single-base mutations in DNA or RNA
Features and Advantages
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Efficient one-tube and extraction-free protocol.
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Ultrafast: Process 96 samples in less than 30 minutes with <10-second Hands-on Time.
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Nuclease Recovered at the end of the reaction thereby can be reused.
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Easy separation of the endonuclease from the reaction.
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Stability of the immobilized nuclease increases.
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Cost-effective: Eliminates columns, filters, laborious, organic reagents, and minimal plasticware required.
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High throughput: Compatible with many different automated liquid handling systems.
Workflow
Formulation:Liquid (Supplied in 50 mM Tris-HCl, pH 8.0, 50% Glycerol.)
Activity:1 μl Magnetic Beads will digest 0.5μg of yeast RNA (Sigma, Catalog # R9001) in 50 mM Tris-HCl, pH 8.0 in 15 minutes at 37 °C.
Shipping:
Shipped at ambient temperature. Upon receipt, store nuclease magnetic Beads at -20°C. Aliquot to avoid repeated freezing and thawing.
PROTOCOL
A. Accessory equipment
Magnetic Rack
Item
Magnetic Rack for centrifuge tube
** Based on sample volume, the user can choose one of the following magnetic Racks
Source
• BcMag™ Rack-2 for holding two individual 1.5 ml centrifuge tubes (Bioclone, Cat. No. MS-01)
• BcMag™ Rack-6 for holding six individual 1.5 ml centrifuge tubes (Bioclone, Cat. No. MS-02)
• BcMag™ Rack-24 for holding twenty-four individual 1.5-2.0 ml centrifuge tubes (Bioclone, Cat. No. MS-03)
• BcMag™ Rack-50 for holding one 50 ml centrifuge tube, one 15 ml centrifuge tube, and four individual 1.5 ml centrifuge tubes (Bioclone, Cat. No. MS-04)
Item
BcMag™ 96-well Plate Magnetic Rack.
Source
• BcMa™ 96-well Plate Magnetic Rack (side-pull) compatible with 96-well PCR plate and 96-well microplate or other compatible Racks (Bioclone, Cat. No. MS-05)
Items
Magnetic Rack for centrifuge tube
** Based on sample volume, the user can choose one of the following magnetic Racks
Source
●
BcMag™ Rack-2 for holding two individual 1.5 ml centrifuge tubes (Bioclone, Cat. No. MS-01)
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BcMag™ Rack-6 for holding six individual 1.5 ml centrifuge tubes (Bioclone, Cat. No. MS-02)
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BcMag™ Rack-24 for holding twenty-four individual 1.5-2.0 ml centrifuge tubes (Bioclone, Cat. No. MS-03)
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BcMag™ Rack-50 for holding one 50 ml centrifuge tube, one 15 ml centrifuge tube, and four individual 1.5 ml centrifuge tubes (Bioclone, Cat. No. MS-04)
BcMag™ 96-well Plate Magnetic Rack
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BcMa™ 96-well Plate Magnetic Rack (side-pull) compatible with 96-well PCR plate and 96-well microplate or other compatible Racks (Bioclone, Cat. No. MS-05)
B. Procedure
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Do not use buffers containing organic solvents.
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Typically, the bead is added directly into any standard buffer at the desired amount of the beads based on the concentration of the RNA (1 μl Magnetic Beads will digest 10μg of yeast RNA).
Working Conditions
TemperatureOptimal Function 60 °CFunctional Range 15 - 70 °C
Inhibitory Action N/ApHOptimal Function 7.6
Functional Range 6.0 - 10.0
Inhibitory Action N/AZn2+Inhibitory Action YesCu2+Optimal Function N/A
Functional Range N/A
Inhibitory Action Yes0-100 mM NaClOptimal Function
Preferentially cleave single-stranded and double-stranded RNA and the RNA strand in RNA-DNA hybrids.
Functional Range N/APrevious slideNext slide
Working Conditions
Conditions
Temperature
pH
Zn2+Cu2+Optimal Function
60 °C
7.6
N/A
Functional Range
15 – 70 °C
6.0 – 10.0
N/A
Inhibitory Action
N/A
N/A
YES
YES
0-100 mM NaCl
Preferentially cleave single-stranded and double-stranded RNA and the RNA strand in RNA-DNA hybrids.
N/A
>300 mM NaCl
Preferentially cleave single-stranded RNA
Guanidine HCl
N/A
N/A
4M + 0.1 M 2-mercaptoethanolSDS
YES1.
Shake the bottle to resuspend the Magnetic beads until it is homogeneous entirely.
IMPORTANT! It is essential to mix the beads before dispensing. Do not allow the beads to sit for more than 2 minutes before dispensing. Resuspend the magnetic beads every 2 minutes.
2.
Add an appropriate amount of the magnetic Beads to a reaction.
3.
Mix the sample with beads for 1-2 minutes by slowly pipetting up and down 20-25 times or Vortex the sample for 2 minutes at 2000 rpm.
4.
Incubate at 37°C with continuous rotation for 15 minutes.
5.
Place the sample plate or tube on the magnetic Rack for 30 seconds or until the solution is clear.
(Option: centrifuge at 3500 rpm for 45 seconds)
6.
Transfer the supernatant to a clean plate /tube while the sample plate remains on the magnetic separation plate. The sample is ready for downstream applications.
Learn More
Instruction Manual
MSDS
DNA & RNA Purification Related Products →General Reference
1.
Molecular Cloning, A Laboratory Manual (3rd ed). Cold Spring Harbor Laboratory Press (Cold Spring Harbor, NY), Volume 1, 1.78-1.79 (2001).
