Blood RNA Purification Kit (Magnetic Bead )

product composition

Product storage

Buffer L9 can be transported at room temperature, please store at 2 ~ 8 ℃ after receipt. Other reagents and articles can be stored at room temperature (0~30°C), and the validity period is two years.

Product description

This product is designed for a 96- well magnetic bead automatic nucleic acid extractor, suitable for isolating and purifying total RNA from 500 µl freshly collected or - 80 ℃ frozen EDTA anticoagulated whole blood or bone marrow . After the anticoagulated whole blood is pretreated by Buffer L9, you only need to add the pretreated blood sample supernatant to the pre-packed plate 1, and the instrument can automatically complete a series of processes such as adsorption, washing and elution of the total blood RNA in the sample. The final blood total RNA is eluted in the plate 5 and can be used immediately for various molecular biology experiments.

Reagents and items to be prepared by the user

1. RNase-free 2 ml centrifuge tube

2. RNase-free pipettes and pipette tips (to avoid contamination between samples, please choose pipette tips with filter elements)

3. Latex gloves, disposable masks and other protective equipment and paper towels

4. Desktop small centrifuge (can be equipped with rotors for centrifuging 1.5ml centrifuge tubes and 2ml centrifuge tubes)

5. Vortexer

Prepare before use

1) Since saliva and skin contain RNase, please wear a mask and latex gloves during the whole process of RNA extraction.

2) Try to use fresh whole blood or bone marrow within 3 hours of isolation for RNA extraction, otherwise the final RNA recovery efficiency will be affected due to RNA degradation. If fresh whole blood cannot be extracted in time for RNA extraction, the whole blood can be lysed in Buffer L9 and frozen at -20 °C or lower ( -70 °C). (refer to step 1 of protocol for details)

Protocol:

1.       Add 1ml of Buffer L9 ito a 2 ml centrifuge tube (centrifuge tube with a cover, prepared by the user), then add 500 μl of whole blood or bone marrow (EDTA anticoagulant), shake vigorously for more than 5 times, and vortex vigorously for 30 seconds to mix well.

* If RNA extraction cannot be performed on freshly obtained whole blood or bone marrow in time, the whole blood lysate can be frozen at -20 °C in this step. The whole blood lysate can be frozen at -20 ℃ for at least two weeks without affecting the extraction efficiency of RNA.

* If the EDTA anticoagulated whole blood frozen at -80 ℃ is used to extract RNA, DO NOT frozen and thawed blood repeatedly. If RNA is extracted from blood samples that have been frozen and thawed more than once, degraded RNA bands will be observed . If the blood sample need to be used repeatedly, please divide the blood sample into small portions and freeze it.

* Buffer L9 is corrosive, please wear protective equipment for operation.

2.       Centrifuge at full speed (≥ 13 000 rpm) for 1 minute.

3.       Tear off the aluminum foil on plate 1, pipette 700 µl of the centrifuged supernatant and transfer it to each well where the reagents have been dispensed .

4.       Put the plate 1 with the sample into the card slot of the instrument, and then discharge the plates 2 to 5 into the card slot of the instrument in sequence, and tear off the aluminum foil on the plates 2 to 5 . Note that Plate 5 needs to be placed in the card slot with the heating module.

5.       Set the operating program of each deep well plate according to the following table:

6.       Take plate 5 out, seal it with parafilm, and store it at -80°C for later use.

* Since the magnetic beads will take away part of the eluted RNA, the collected RNA is about 60 μl.