Introduction

Hipure Stool RNA Kit is specially designed for stool RNA extraction. This kit is suitable for extracting high-purity microbial or host cell RNA from ≤0.1g stool samples. The kit adopts silica gel column purification technology and original solution system, which can effectively remove humic acid and other inhibitory factors in stool samples. The purified RNA can be directly used in RT-PCR, Northern hybridization and other experiments.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from 100-150mg stool sample
Applications	RT-PCR, Northern hybridization and other experiments
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Stool
Sample amount	100-150 mg
Elution volume	≥30μl
Time per run	≤50 minutes
Liquid carrying volume per column	100µg
Binding yield of column	800µl

Principle

The HiPure silica gel column uses a high binding ability glass fiber filter membrane as the substrate. Under the condition of high concentration of ionizing agent (such as Guanidinium chloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bonding and electrostatic and other physical factors, while protein or other impurities are not adsorbed and removed. The filter membrane that has adsorbed nucleic acids is washed to remove proteins and salts. Finally, low salt buffer solution (such as Buffer TE) or water can be used to wash out the nucleic acids adsorbed on the filter membrane. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.

The stool samples are homogenized in the lysis solution, further lysed in a high-temperature water bath, and RNA is released into the lysis solution. Chloroform extraction removes genomic DNA and impurities, transfer the supernatant to an alcohol free binding solution, purify RNA through a column, and finally elute RNA with RNase Free Water. The purified RNA can be directly used for experiments such as PCR, Southern hybridization, and enzyme digestion.

Advantages

• High purity – unique adsorbent for more efficient removal of inhibitors
• High concentration – maximum extraction of total RNA from stool samples
• Sensitive – RNA can be purified at the level of PG

Kit Contents

Contents	R418502	R418503
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	50	250
Glass Beads (0.1~0.6mm)	30 g	150 g
Buffer SPL	30 ml	140 ml
Buffer PHC	30 ml	140 ml
Buffer GRP	60 ml	250 ml
Buffer RW1	50 ml	250 ml
Buffer RW2 *	20 ml	2 x 50 ml
RNase Free Water	15 ml	30 ml

Storage and Stability

The kit components can be stored at room temperature (15–25°C) and are stable for 18 months under these conditions. At low temperatures, Buffer SPL may form precipitates, dissolve it by 55°C water bath. After receiving the product, Buffer PHC should be stored at 2-8°C.

Hipure Stool RNA Kit is specially designed for stool RNA extraction. This kit is suitable for extracting high-purity microbial or host cell RNA from ≤0.1g stool samples. The kit adopts silica gel column purification technology and original solution system, which can effectively remove humic acid and other inhibitory factors in stool samples. The purified RNA can be directly used in RT-PCR, Northern hybridization and other experiments.

t-Boc-aminooxy-PEG2-propargyl is a click chemistry crosslinker. The propargyl group is reactive with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry to yield a stable triazole linkage. t-Boc-aminooxy can be deprotected under mild acidic conditions. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

t-Boc-aminooxy-PEG2-propargyl is a click chemistry crosslinker. The propargyl group is reactive with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry to yield a stable triazole linkage. t-Boc-aminooxy can be deprotected under mild acidic conditions. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

About

A universal PCR master mix for allele-specific PCR assays. Precision fluorescent signal generation with consistently high performance at any reaction volume.

PACE Genotyping Master Mix uses a novel, universal, fluorescent reporting cassette to produce machine-readable fluorescent signals corresponding to genotypes. PACE compatible genotyping assays are comprised of two competitive allele specific forward primers (which differ in their terminal 3’ bases and unique 5’ tail sequences) and a common, reverse primer.  PACE Genotyping Master Mix is supplied at 2x concentration and with ROX normalising dye at a range of levels to ensure compatibility with your qPCR instrument or reader.

Genotyping assay designs are available from 3CR Bioscience through our free PACE assay-design service; once designed, users can purchase assay primers independently or through 3CR Bioscience using our partial or full-assay validation service. PACE Genotyping Master Mix is also compatible with KASP™ and Amplifluor® marker assays.

REQUIRED COMPONENTS

• qPCR machine or Thermocycler + Fluorescent plate reader
• PCR plate or equivalent and appropriate optically clear seal
• Template DNA
• PCR-grade water
• Genotyping assays

qPCR machine or Thermocycler + Fluorescent plate reader

PCR plate or equivalent and appropriate optically clear seal

Template DNA

PCR-grade water

Genotyping assays