Einführung

• Purpose: Extraction of stool RNA.
• Sample Compatibility: Suitable for extracting high-purity microbial or host cell RNA from ≤0.1g stool samples.
• Technology: 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.
• Anwendungen: The purified RNA can be directly used in RT-PCR, Northern hybridization, and other experiments.

Spezifikationen

Prinzip

• Principle of HiPure Silica Gel Column: Utilizes a high binding ability glass fiber filter membrane as the substrate. Under high concentrations of ionizing agents (z.B., Guanidinium chloride or Guanidin isothiocyanate), nucleic acids are adsorbed through hydrogen bonding and electrostatic interactions, while proteins and other impurities are not adsorbed and removed. Anschließend, the filter membrane with adsorbed nucleic acids is washed to remove proteins and salts. Finally, low salt buffer solution (z.B., Buffer TE) or water is used to elute the purified nucleic acids, resulting in high purity suitable for downstream experiments.
• Probenverarbeitung: 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. Finally, RNA is eluted using RNase Free Water.
• Anwendungen: The purified RNA is suitable for various downstream experiments such as PCR, Southern hybridization, and enzyme digestion.

Vorteile

• Hohe Reinheit: 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.

Inhalt des Kits

Lagerung und Stabilität

• Storage Conditions: Kit components can be stored at room temperature (15–25°C) and remain stable for 18 Monate unter diesen Bedingungen.
• 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.