Description
mRNA capture beads
GenieSeq mRNA Capture Beads are oligo(dT) magnetic beads for selective enrichment of poly(A) mRNA from total RNA ahead of RNA-seq library preparation. They provide efficient, reproducible mRNA capture with low rRNA carryover and are compatible with manual and automated workflows.
The beads are 1 μm oligo(dT)-coupled paramagnetic microspheres that isolate poly(A)+ RNA from purified total RNA. Magnetic separation recovers intact mRNA in small volumes without precipitation, and the whole workflow takes about 1 h. The kit handles 0.01–12.5 μg of high-integrity total RNA (RIN ≥7) from a wide range of species, including animals, plants and fungi. The enriched mRNA can go straight into reverse transcription or RNA-seq library preparation. For research use only.
Key Features
| High purity | Specific mRNA enrichment gives higher purity with reduced rRNA residue. | |
| Wide compatibility | Works with animal, plant and fungal samples and inputs from 10 ng to 12.5 μg. | |
| Fast workflow | Isolates intact mRNA in about 1 h with no precipitation step. | |
| Library-prep ready | Eluted mRNA can be used directly for reverse transcription or RNA-seq library preparation. |
Specifications
| Technology | 1 μm oligo(dT)-coupled paramagnetic microspheres |
| Sample | Purified total RNA, 0.01–12.5 μg (RIN ≥7) |
| Species | Animals, plants and fungi |
| Workflow time | About 1 h |
| Applications | Poly(A)+ mRNA isolation for RNA-seq library preparation and reverse transcription |
| Shelf life | 2 years |
Components
| Components | 24 rxns | 96 rxns |
| Beads Binding Buffer 2.0 | 1.2 ml | 4.8 ml |
| mRNA Capture Beads 2.0 | 1.2 ml | 4.8 ml |
| Beads Wash Buffer 2.0 | 9.6 ml | 38.4 ml |
| Tris Buffer 2.0 | 1.2 ml | 4.8 ml |
| Nuclease-free ddH2O | 1 ml | 4 × 1 ml |
Not supplied: low-binding nuclease-free PCR tubes and pipette tips, a PCR instrument and a magnetic rack.
Storage
Store at 2 ~ 8°C.
Perform all steps at room temperature to ensure capture efficiency.
- Prepare reagents and sample
- Equilibrate the reagents from 2–8°C storage to room temperature.
- Dilute 0.01–12.5 μg of total RNA to 50 μl with nuclease-free ddH2O in a nuclease-free PCR tube and keep on ice.
- First capture
- Mix the mRNA Capture Beads 2.0 by inversion. Add 50 μl of beads to the RNA and pipette up and down 6 times.
- In a PCR instrument, incubate at 65°C for 5 min, then 25°C for 5 min, and hold at 4°C to bind mRNA to the beads.
- Place on a magnetic rack for 5 min. When the solution is clear, carefully discard the supernatant.
- Wash
- Remove from the rack, add 200 μl of Beads Wash Buffer 2.0 and pipette up and down 6 times.
- Place on the rack for 5 min. When clear, carefully discard the supernatant.
- Elute
- Remove from the rack, add 50 μl of Tris Buffer 2.0 and pipette up and down 6 times to resuspend the beads.
- In a PCR instrument, incubate at 80°C for 2 min and hold at 25°C to elute the mRNA.
- Second capture
- Add 50 μl of Beads Binding Buffer 2.0 and pipette up and down 6 times. Incubate at room temperature for 5 min.
- Place on the rack for 5 min. When clear, carefully discard the supernatant.
- Wash
- Remove from the rack, add 200 μl of Beads Wash Buffer 2.0 and pipette up and down 6 times.
- Place on the rack for 5 min. When clear, carefully discard the supernatant.
- Final elution for downstream use
- For reverse transcription: remove from the rack, add 10.5 μl of nuclease-free ddH2O and pipette up and down 6 times. Incubate at 80°C for 2 min, then immediately place on the rack for 5 min. When clear, transfer 8 μl of supernatant to a new nuclease-free PCR tube.
- For RNA library preparation: add the volume of Frag/Prime Buffer specified in your RNA-seq library prep kit instructions and continue library preparation.
- Use or store
- Place on ice and use immediately for NGS library preparation or other analyses (recommended), or store at -85 ~ -65°C.
Q1: How much total RNA can I use?
- The beads isolate poly(A)+ RNA from 0.01–12.5 μg of high-integrity total RNA (RIN ≥7), diluted to 50 μl.
Q2: Can I use partially degraded RNA?
- Use high-integrity RNA (RIN ≥7). Degraded total RNA may lead to partial loss of poly(A)+ RNA.
Q3: What do I need to supply myself?
- Low-binding nuclease-free PCR tubes and pipette tips, a PCR instrument and a magnetic rack.
Q4: How should I store the purified mRNA?
- Use it immediately for library preparation or other analyses if possible; otherwise store it at -85 ~ -65°C.
