소개
- 목적: Extraction of stool RNA.
- 샘플 호환성: Suitable for extracting high-purity microbial or host cell RNA from ≤0.1g stool samples.
- 기술: Utilizes silica gel column purification technology and an original solution system.
- Key Feature: Effectively removes humic acid and other inhibitory factors present in stool samples.
- 응용: The purified RNA can be directly used in RT-PCR, Northern hybridization, and other experiments.
명세서
특징 | 명세서 |
주요 기능 | Isolation total RNA from 100-150mg stool sample |
응용 | RT-PCR, Northern hybridization, and other experiments |
정제방법 | 미니 스핀 컬럼 |
정제기술 | 실리카 기술 |
가공방법 | 수동 (원심분리 또는 진공) |
샘플 유형 | Stool |
샘플량 | 100-150 mg |
용출량 | ≥30μl |
실행당 시간 | 50분 이하 |
컬럼당 액체 운반량 | 100µg |
컬럼의 결합 수율 | 800μl |
원칙
- Principle of HiPure Silica Gel Column: Utilizes a high binding ability glass fiber filter membrane as the substrate. Under high concentrations of ionizing agents (예를 들어, Guanidinium chloride or 구아니딘 isothiocyanate), nucleic acids are adsorbed through hydrogen bonding and electrostatic interactions, while proteins and other impurities are not adsorbed and removed. Subsequently, the filter membrane with adsorbed nucleic acids is washed to remove proteins and salts. 마지막으로, low salt buffer solution (예를 들어, Buffer TE) or water is used to elute the purified nucleic acids, resulting in high purity suitable for downstream experiments.
- 샘플 처리: Stool samples are homogenized in the lysis solution, then further lysed in a high-temperature water bath to release RNA into the solution. Chloroform extraction is employed to remove genomic DNA and impurities. The supernatant is transferred to an alcohol-free binding solution, followed by RNA purification through a column. 마지막으로, RNA is eluted using RNase Free Water.
- 응용: The purified RNA is suitable for various downstream experiments such as PCR, Southern hybridization, and enzyme digestion.
장점
- 고순도: Utilizes a unique adsorbent for more efficient removal of inhibitors, ensuring high purity of extracted RNA.
- High Concentration: Achieves maximum extraction of total RNA from stool samples due to its high concentration, maximizing RNA yield.
- Sensitive: Capable of purifying RNA at the level of picograms (PG), ensuring sensitivity in downstream applications.
키트 내용물
내용물 | R418502 | R418503 |
정화 시간 | 50 준비 | 250 준비 |
HiPure RNA Mini Columns | 50 | 250 |
2ml 수집 튜브 | 50 | 250 |
Glass Beads (0.1~0.6mm) | 30 g | 150 g |
Buffer SPL | 30 밀리리터 | 140 밀리리터 |
Buffer PHC | 30 밀리리터 | 140 밀리리터 |
Buffer GRP | 60 밀리리터 | 250 밀리리터 |
Buffer RW1 | 50 밀리리터 | 250 밀리리터 |
Buffer RW2 * | 20 밀리리터 | 2 엑스 50 밀리리터 |
RNase Free Water | 15 밀리리터 | 30 밀리리터 |
보관 및 안정성
- 보관 조건: Kit components can be stored at room temperature (15-25°C) and remain stable for 18 이런 조건에서 몇 달 동안.
- Buffer SPL: May form precipitates at low temperatures. Dissolve precipitates by placing the Buffer SPL in a 55°C water bath.
- Buffer PHC: After receiving the product, store Buffer PHC at 2-8°C.
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