| Field | Specification |
|---|---|
| Mfr No | |
| Product Type | |
| Shipping | |
| Storage |
Description
This product is suitable for total RNA extraction from cells and animal tissues (such as liver, kidney, spleen, brain, mouse tail, etc.). Utilizing a unique magnetic bead system and a carefully optimized buffer system, it effectively captures released nucleic acids. The extracted RNA has high purity and stable quality, and is suitable for downstream applications such as RT-PCR, Northern Blot, and in vitro translation. When used in conjunction with automated nucleic acid extraction instruments, this kit enables high-throughput nucleic acid isolation.
Features
High yield and purity of RNA, free of proteins, DNA, and contaminants
Compatible with a variety of tissue types and cultured cells
Fast and reproducible protocol, preserving RNA integrity
Suitable for both low- and high-throughput applications
Components
|
Components No. |
Name |
18600ES50 |
18600ES60 |
|
18600-A |
Lysis Buffer |
23 mL |
45 mL |
|
18600-B |
Wash Buffer A* |
44 mL |
44 mL × 2 |
|
18600-C |
Wash Buffer B* |
18 mL |
18 mL × 2 |
|
18600-D |
Elution Buffer |
5 mL |
10 mL |
|
18600-E |
Magnetic Bead Suspension |
1 mL × 2 |
1 mL × 4 |
[Note]: For 18600ES50: Prior to use, add 44 mL of absolute ethanol to each bottle of Wash Buffer A (18600-B), and 72 mL of absolute ethanol to each bottle of Wash Buffer B (18600-C).
For 18600ES60: Prior to use, add 44 mL of absolute ethanol to each bottle of Wash Buffer A (18600-B), and 72 mL of absolute ethanol to each bottle of Wash Buffer B (18600-C).
Storage
This product should be stored at room temperature for 1 year.
Application
Tissue/Cell Total RNA Extraction; High-purity RNA Extraction; Gene Expression Analysis.
Instructions
Prepare lysis buffer according to sample volume. Add β-mercaptoethanol or DTT to the lysis buffer to a final concentration of 1%. Use freshly prepared lysis buffer immediately.
Sample Pre-treatment
A. Tissue Samples
Weigh 5–20 mg of tissue sample and add 450 μL of prepared lysis buffer. Homogenize thoroughly. Centrifuge at 12,000 rpm for 2 min and collect the supernatant to obtain the sample lysate.
B. Cell Samples
For up to 500,000 cells: Pellet cells by centrifugation, remove supernatant, and resuspend in 450 μL of prepared lysis buffer. Vortex or pipette to fully lyse cells. Incubate at room temperature for 5 min to obtain the sample lysate.
For 500,000–1,000,000 cells: Pellet cells by centrifugation, remove supernatant, and resuspend in 500 μL of prepared lysis buffer. Vortex or pipette to fully lyse cells. Incubate at room temperature for 5 min to obtain the sample lysate.
Manual RNA Extraction Protocol
1.Transfer 450 μL of sample lysate to a 1.5 mL microcentrifuge tube. Add 400 μL of isopropanol and 20 μL of Magnetic Bead Suspension (for 500,000–1,000,000 cells, use 500 μL lysate, 400 μL isopropanol, and 40 μL Magnetic Bead Suspension). Vortex thoroughly to fully disperse the magnetic beads. Place the tube on a rotating mixer and incubate for 10 min.
[Note]: The Magnetic Bead Suspension must be thoroughly mixed before use.
2.After incubation, briefly centrifuge the tube and place it on a magnetic stand. Wait until all magnetic beads are completely pulled to the side of the tube. Carefully remove and discard the supernatant.
3.Add 800 μL of Wash Buffer A (ensure ethanol has been added) to the tube. Vortex vigorously for 1 min to resuspend the beads. Place the tube back on the magnetic stand until the beads are fully captured. Carefully remove and discard the supernatant.
4.(Optional: DNase I treatment for DNA removal) Add 6 μL of DNase I and 74 μL of RDD Buffer. Briefly vortex to disperse the beads. Incubate at room temperature for 15 min, vortexing every 5 min.
5.Repeat Step 3.
6.Add 800 μL of Wash Buffer B (ensure ethanol has been added). Vortex vigorously for 1 min. Place the tube on the magnetic stand until the beads are fully captured. Carefully remove and discard the supernatant.
7.Repeat Step 6.
8.Open the tube lid and air-dry at room temperature or 37°C until the surface of the magnetic bead pellet just begins to crack. Avoid over-drying.
9.Add 70 μL of Elution Buffer. Vortex thoroughly or pipette up and down to fully resuspend the beads. Incubate at 56°C for 5 min, vortexing several times during incubation.
10.Place the tube on the magnetic stand until the beads are fully captured. Transfer the supernatant to a RNase-free microcentrifuge tube, taking care not to transfer any magnetic beads. Store the RNA solution at -80°C.
Documents:
Safety Data Sheet
Manuals
