The Kit provides fast purification of high-quality RNA from paxigene Blood RNA Tube using silica-membrane spin columns. RNA purified using the HiPure System is ready for applications such as RT-PCR, 노던 블로팅, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
세부
명세서
| 특징 | 명세서 |
| 주요 기능 | Isolation of total RNA from Paxgene RNA Tubes |
| 응용 | RT-PCR, qPCR, Northern hybridization, NGS, nucleic acid protection, in vitro translation |
| 정제방법 | 미니 스핀 컬럼 |
| 정제기술 | 실리카 기술, DNA filtration technology |
| 가공방법 | 수동 (원심분리 또는 진공) |
| 샘플 유형 | Blood in the preservation tube |
| 샘플량 | 2.5밀리리터 |
| 용출량 | ≥30μl |
| 컬럼당 액체 운반량 | 800μl |
| 컬럼의 결합 수율 | 100μg |
원칙
The Kit is for the purification of total RNA from 2.5 ml human whole blood collected in a PAXgene Blood RNA Tube. Purification begins with a centrifugation step to pellet nucleic acids in the PAXgene Blood RNA Tube. The pellet is washed and resuspended, and incubated in optimized buffers together with proteinase K. DNA wash The lysate is passed through a DNA Mini column. Ethanol is added to adjust binding conditions, and the lysate is applied to a column.RNA is selectively bound to the silica membrane as contaminants pass through. The remaining contaminants are removed in several efficient wash steps. Between the first and second wash steps, the membrane is treated with DNase I to remove trace amounts of bound DNA. After the wash steps, RNA is eluted in elution buffer and heat-denatured.
장점
- High quality – high purity total RNA can be directly used in various sensitive downstream applications
- 빠른 – isolation of several samples can be completed in 40 minutes by using the column purification method
- Safety – no phenol-chloroform extraction is required
- Sensitive – direct lysis of blood, 혈장, and other samples without separation of leukocytes
키트 내용물
| 내용물 | R416802 | R416803 |
| 정화 시간 | 10 준비 | 50 준비 |
| HiPure RNA Mini Columns I | 10 | 50 |
| gDNA Filter Mini Columns | 10 | 50 |
| 2ml 수집 튜브 | 30 | 150 |
| RNase Free Water | 60 밀리리터 | 250 밀리리터 |
| Buffer MBR1 | 10 밀리리터 | 30 밀리리터 |
| Buffer MBR2 | 5 밀리리터 | 15 밀리리터 |
| Buffer RW1 | 15 밀리리터 | 60 밀리리터 |
| Buffer RW2* | 6 밀리리터 | 20 밀리리터 |
| 단백질분해효소 K | 12 mg | 50 mg |
| 프로테아제 용해 완충액 | 1.8 밀리리터 | 5 밀리리터 |
| DNase I | 120 μl | 600 μl |
| DNase Buffer | 6 밀리리터 | 30 밀리리터 |
보관 및 안정성
Proteinase K should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. 하지만, 단기 보관 (DNase I up to 1 주, Proteinase K up to 8 주) 실온에서 (15-25°C) 성능에 영향을 미치지 않습니다. 나머지 키트 구성품은 실온에서 보관 가능 (15-25°C) 그리고 적어도 안정적이다 18 이런 조건에서 몇 달 동안.
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