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AllPrep PowerFecal DNA/RNA Kit

For simultaneous purification of genomic DNA and total RNA from stool samples
  • Efficient removal of inhibitors through Inhibitor Removal Technology (IRT)
  • Separates RNA and DNA in different eluates for easier downstream analysis
  • Optimized lysis of microbial cells for increased DNA and RNA yields
  • Enables comprehensive and comparative metagenomic and metatranscriptomic analysis
For reliable comparison of genomic and transcriptomic data, purification of DNA and RNA from the same sample is essential. The AllPrep PowerFecal DNA/RNA Kit protocol enables the simultaneous isolation of DNA and RNA in separate eluates from one sample. The kit is optimized for use in metagenomic and metatranscriptomic analyses using next-generation sequencing (NGS).
Cat No./ID: 80244
AllPrep PowerFecal DNA/RNA Kit
For 50 preps: 50 AllPrep DNA MinElute spin columns, 50 RNeasy Mini spin columns, Collection Tubes, Elution Tubes and Buffers
The AllPrep PowerFecal DNA/RNA Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
0
Inhibitor removal.
DNA and RNA was prepared from eight different human stool samples (200 mg each) using a standard protocol (AllPrep DNA/RNA Mini Kit, without IRT) and with the AllPrep DNA/RNA PowerFecal Kit (with IRT). DNA and RNA purified using the AllPrep PowerFecal DNA/RNA Kit shows no inhibition in a PCR assay.
1
OTU analysis with different stool samples.
DNA from human, mouse, sheep and horse stool samples (200 mg each) was prepared using the AllPrep DNA/RNA Power Fecal Kit and sheared to 300 bp fragments. Paired-end NGS libraries were prepared, quantified and sequenced. BWA-MEM Version 0.7.10 was used to map DNA reads against the human reference genome Hg19, resulting in a “human DNA-free” data set. Unmapped reads were subsequently mapped against a collection of bacterial genomes.
2
The AllPrep PowerFecal DNA/RNA Kit procedure.
Performance
The AllPrep PowerFecal DNA/RNA Kit efficiently purifies DNA and RNA from stool samples that are inherently rich in PCR inhibitors. Removing such inhibitors is crucial to obtain a good signal in PCR or NGS analysis. The AllPrep PowerFecal DNA/RNA Kit comes with the patented Inhibitor Removal Technology (IRT) ensuring complete removal of inhibitory substances from digested food, heme from lysed red blood cells in stool and other PCR inhibitors (see figure “Inhibitor removal”). The AllPrep PowerFecal DNA/RNA Kit uses improved Microbial Lysis Tubes to ensure high yields of both DNA and RNA from stool samples. The DNA and RNA are suitable for NGS analysis such as microbial operational taxonomic unit (OTU) analysis or comparative metagenomic and metatranscriptomic studies (see figure “OTU analysis with different stool samples”).
Principle
The AllPrep PowerFecal DNA/RNA Kit is designed for simultaneous purification of genomic DNA and total RNA from stool samples. Pure DNA and RNA are obtained from the entire sample, in contrast to other procedures where the biological sample is divided into two before being processed separately. Simply dividing a sample in half for separate DNA and RNA purification procedures, results in the purification of DNA and RNA from different populations of cells, introducing a bias in any downstream analysis.
Procedure
A simple workflow allows the purification of high-quality DNA and RNA from the same stool sample (see flowchart “AllPrep PowerFecal DNA/RNA Kit procedure”).
Stool samples are first homogenized followed by a combined chemical and mechanical lysis using prefilled lysis tubes. Inhibitors commonly present in stool samples are then removed before isolation of nucleic acids. Genomic DNA is first immobilized on a silica membrane while RNA molecules pass through. This flow-through is applied to a different column to specifically bind RNA. The bound DNA and RNA, respectively, are further washed and eluted to yield the different nucleic acids in separate eluates. Optional on-column DNase and RNase digestion can further eliminate any traces of cross-contaminating nucleic acids, if required.
Applications
The AllPrep PowerFecal DNA/RNA Kit is optimized to remove inhibitors and lyse microbial cells efficiently by using the patented IRT technology and bead-beating steps, respectively. The DNA and RNA obtained with the AllPrep PowerFecal DNA/RNA Kit are suitable for use in NGS enabling applications such as 16S rDNA profiling, metagenomic shotgun sequencing and metatranscriptomic profiling. As DNA and RNA are isolated in parallel from the same sample, the AllPrep PowerFecal DNA/RNA Kit is ideal for comparative metagenomic and metatranscriptomic analysis.
Features
Specifications
Applications Comparative metagenomic and metatranscriptomic analysis using NGS, PCR and qPCR
Elution volume 60 µl
Format Spin column
Main sample type Stool
Number of preps per run 50
Processing Manual (centrifugation)
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein Total RNA and genomic DNA
Sample amount 100–200 mg
Technology Silica technology
Time per run or per prep 45 min
Yield Varies
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