Blood Total RNA Extraction Kit (Genomic DNA Removal)

1. 試劑盒的組成

2. 貯存

該試劑盒應在室溫下保存 (15-25℃) in a dry environment and can be preserved for 12 月.

3. 試劑盒使用說明

3.1 該試劑盒旨在用於分子生物學研究目的，不應用於疾病診斷或治療.

3.2 套件中的某些成分含有刺激物; 建議採取必要的預防措施 (例如穿著防護衣和護目鏡).

3.3 使用該套件需要額外的設備，例如高速離心機, 水浴 (金屬浴), 渦旋混合器, 無水乙醇, 液態氮, 氯仿, 無菌去離子水, 和EP管.

4. 試劑盒簡介

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 毫升, 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 (包括核RNA和細胞質RNA) 來自體內的血液 2 小時. The purified RNA can be directly used for applications such as RT-聚合酶鍊式反應, 北方印跡, ETC. 整個純化過程不需要苯酚氯仿等有毒試劑, making the RNA purification kit well-suited for various other sample types.

5. 實驗原理和程序

6. 提取過程

開始實驗前的注意事項:

A. 使用前, 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. EDTA是否影響後續實驗, it is recommended to use sterile deionized water as a substitute for the Elution Buffer.

1. 樣品加工: Take 200μL of blood into an EP tube, 添加 80μL of Red Blood Cell Lysis Buffer 10x, and mix thoroughly by pipetting.
2. 添加 500μL RNA Extraction Buffer 1 加, vigorously shake to mix, centrifuge at 12000rpm for 3 分分鐘.
3. Transfer the supernatant to a DNA純化 柱子, centrifuge at 12000rpm for 1 分分鐘.
4. 添加 250μL of ethanol to the supernatant, pipette and mix, transfer the liquid to the RNA purification column, centrifuge at 12000rpm for 30 秒, 棄去上清液.
5. 添加 600μL Inhibitor Removal Buffer 至RNA純化柱, centrifuge at 12000rpm for 30 秒, 棄去上清液.
6. 添加 600μL Wash Buffer 1 至RNA純化柱, centrifuge at 12000rpm for 30 秒, 棄去上清液.

(筆記: Confirm that ethanol has been added to Wash Buffer 1. Ethanol presence significantly affects subsequent experiments. 所以, membrane drying is crucial. 離心後, ensure no ethanol remains before elution, then discard the waste and collection tube. 使用洗滌緩衝液後 1, the membrane on the RNA purification column should have a slight color. 離心後, carefully remove the RNA purification column, ensuring it does not touch the collection tube to prevent ethanol interference.)

7. 重複步驟 6.
8. Centrifuge the empty tube at 12000rpm for 2 分分鐘 (to evaporate residual ethanol).
9. 將 RNA 純化管柱放入新的離心管中, drip 100μL Elution Buffer onto the membrane, 室溫孵育 5 分分鐘 (15℃~25℃), centrifuge at 12000rpm for 1 分分鐘.

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