Blood Total RNA Extraction Kit (Genomic DNA Removal)

1. Components of the reagent kit

2. Storage

This reagent kit should be stored at room temperature (15-25℃) in a dry environment and can be preserved for 12 months.

3. Instructions for Using the Reagent Kit

3.1 This kit is intended for molecular biology research purposes and should not be used for disease diagnosis or treatment.

3.2 Some components in the kit contain irritants; it is advisable to take necessary precautions (such as wearing protective clothing and goggles).

3.3 The use of this kit requires additional equipment such as a high-speed centrifuge, water bath (metal bath), vortex mixer, anhydrous ethanol, liquid nitrogen, chloroform, sterile deionized water, and EP tubes.

4.  Introduction to the Reagent Kit

The RNA purification kit provides a fast and efficient method for purifying RNA from blood and other fluid tissues. It is suitable for most biological fluids and tissues. This RNA purification kit can be applied to blood samples exceeding 0.5 ml, and through DNA removal technology, the extracted RNA is virtually free of genomic DNA.

The RNA Fast Purification Kit allows for the extraction of total RNA (including nuclear RNA and cytoplasmic RNA) from blood within 2 hours. The purified RNA can be directly used for applications such as RT-PCR, Northern blotting, etc. The entire purification process does not require toxic reagents such as phenol-chloroform, making the RNA purification kit well-suited for various other sample types.

5. Experimental Principles and Procedures

6. Extraction Process

Precautions before starting the experiment:

A. Before use, add the specified amount of ethanol to Wash Buffer 1 as indicated on the reagent bottle label, and mark a check on the label to indicate the addition of ethanol.

B.Elution Buffer is a 0.1x TE solution containing a minimal amount of EDTA. If EDTA affects subsequent experiments, it is recommended to use sterile deionized water as a substitute for the Elution Buffer.

1. Sample Processing: Take 200μL of blood into an EP tube, add 80μL of Red Blood Cell Lysis Buffer 10x, and mix thoroughly by pipetting.
2. Add 500μL RNA Extraction Buffer 1 plus, vigorously shake to mix, centrifuge at 12000rpm for 3 minutes.
3. Transfer the supernatant to a DNA purification column, centrifuge at 12000rpm for 1 minute.
4. Add 250μL of ethanol to the supernatant, pipette and mix, transfer the liquid to the RNA purification column, centrifuge at 12000rpm for 30 seconds, discard the supernatant.
5. Add 600μL Inhibitor Removal Buffer to the RNA purification column, centrifuge at 12000rpm for 30 seconds, discard the supernatant.
6. Add 600μL Wash Buffer 1 to the RNA purification column, centrifuge at 12000rpm for 30 seconds, discard the supernatant.

(Note: Confirm that ethanol has been added to Wash Buffer 1. Ethanol presence significantly affects subsequent experiments. Therefore, membrane drying is crucial. After centrifugation, ensure no ethanol remains before elution, then discard the waste and collection tube. After using Wash Buffer 1, the membrane on the RNA purification column should have a slight color. After centrifugation, carefully remove the RNA purification column, ensuring it does not touch the collection tube to prevent ethanol interference.)

7. Repeat step 6.
8. Centrifuge the empty tube at 12000rpm for 2 minutes (to evaporate residual ethanol).
9. Place the RNA purification column in a new centrifuge tube, drip 100μL Elution Buffer onto the membrane, incubate at room temperature for 5 minutes (15℃~25℃), centrifuge at 12000rpm for 1 minute.

(Note: Eluting RNA with 50μL Elution Buffer can increase RNA concentration but reduce total RNA yield.)