Product Description
Product Name MagPure Blood RNA Kit
Product specifications 48 Preps/Kit
Introduction
This product supplies a simple and rapid extraction of total RNA from Blood, buffy coat, bone marrow, Cell suspension and other body fluids. The kit is based on superparamagnetic particles purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction process is only 60 minutes. Purified RNA is ready for downstream applications such as RT-PCR, virus RNA testing and so on. MagPure RNA Kits buffers can be used for both manual extraction process and automatic nucleic acid extraction machines. This Kits is suitable for extracting RNA from ≤ 5x106 cells suspension, 200~300 µl Whole Blood, 200µl buffy coat, 100µl bone marrow.
Main Composition
| Cat.No. |
R661101 |
| Purification times |
48 Preps |
| MagPure RNA Particles |
1.7 ml |
| Proteinase K |
24 mg |
| Protease Dissolve Buffer |
1.8 ml |
| DNase I |
600 µl |
| DNase Buffer |
30 ml |
| Buffer MLB |
40 ml |
| Buffer MCB |
9 ml |
| Buffer MW1 |
22 ml |
| Buffer MW2 |
20 ml |
| RNase Free Water |
10 ml |
Storage conditions and validity
This product can be stored at room temperature (15~25oC) for 12 months. MagPure RNA Particles, Proteinase K, DNase I are shipped at room temperature. After received, store DNase I at -20ºC and store MagPure RNA Particles and proteinase K at 2~8ºC.
Materials and Equipment to be Supplied by User
1.100% ethanol
2.Dilute Buffer MW1 and MW2 with 100% ethanol and store at room temperature
3.Dilute Buffer MCB with store at room temperature
4.Dissolve the Proteinase K with protease Dissolve Buffer to the Proteinase K and store at -20~8ºC
KingFisher or similar Extractor isolation:
1.Add the Reagents/sample to the well of f the deep well plate according to the table below.
| Name of the Plate |
Pre-loaded reagents |
Addition before use |
| Sample plate |
- Pipet 20µl Proteinase K and 30µl MagPure RNA Particles
- Pipet 150~300µl of the sample into the well of plate:
- 200~300 µl whole blood, plasma, serum, body fluids
- 200µl buffy coat or 150µl bone marrow
- up to 5 x 106 lymphocytes or Culture Cells in 200µl PBS.
- Add 600µl Buffer MLB
|
| Wash Plate 1 |
600µl Buffer MW1, Put in 96 magnetic Tip |
| DNase |
290µl DNase Buffer and 10µl DNase I |
| Wash Plate 2 |
600µl Buffer MW2 |
| Wash Plate 3 |
600µl Buffer MW2 |
| Elution plate |
60µl RNase Free Water |
2.Place a 96 tip comb for deep well magnets on Wash Plate 1.
3.Start the R6611_Flex protocol with the KingFisher Flex 96 and load the plates.
4.Add 450µl Buffer MCB to the DNase plate during the dispense step.
5.Place the DNase plate back into the instrument and press Start. After the pause, the protocol will continue to the end.
6.After the run is completed, remove the plates and store the purified total RNA.
Troubleshooting Guide
A:Low RNA yields
1.Incomplete resuspension of MagPure Particles: Resuspend the MagPure Particles by vortexing before use.
2.Loss of MagPure Particles during procedure: Be careful not to remove the MagPure Particles during the procedure.
3.MagPure Particles ot resuspended during binding: Vortex vigorously for 2 minutes after addition of Buffer MCB.
4.Eluate contains residual ethanol: Ensure that the wash flow-through is drained from the collection tube and that the column is then centrifuged at >12,000 x g for 1min.
B:Low A260/A280 value
1.Water used to dilute RNA for A260/A280 measurement: Use 10 mm Tris·Cl, pH 7.5, not RNAse-free water, to dilute the sample before measuring purity..
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