Xetra Bio Solution

For 50 DNA preps: 50 QIAamp UCP Mini Columns, 50 Pathogen Lysis Tubes L, buffers, reagents, Collection Tubes (2 ml) The QIAamp DNA Microbiome Kit is a dedicated solution for the purification and enrichment of bacterial microbiome DNA from swabs and body fluids. Effective depletion of host DNA during the purification process maximizes bacterial DNA coverage in next generation sequencing analysis and allows highly sensitive 16S rDNA-based microbiome analysis and whole metagenome shotgun sequencing studies. Effective host DNA depletion for deeper microbiome analysis

Swabs and body fluids contain DNA from host human or animal cells as well as microbial cells. In metagenomic studies, total DNA is standardly isolated and analyzed, however, host human or animal DNA greatly outweighs the microbial DNA, which can hamper microbiome analysis. In fact, a key finding of the whole metagenome shotgun sequencing of different sample types conducted by the Human Microbiome Project was that up to 99% of the sequencing reads corresponded to the human genome, so that at most 1% were of microbial nature. In contrast to 16S rDNA sequencing, whole metagenome shotgun sequencing can add valuable insight to microbiome studies, such as the presence of virulence factors, antibiotic resistance, or metabolic networks. Thus, maximized coverage of microbial reads greatly enhances analysis power.
Removal of host DNA increases the coverage of microbial reads in sequencing experiments. The QIAamp DNA Microbiome Kit efficiently removes host DNA by differential lysis of host cells and subsequent enzymatic digestion of host DNA. Then, using a combination of mechanical and chemical lysis, intact cells are efficiently lysed and the released bacterial DNA is purified using proven QIAamp chemistry and decontaminated QIAamp UCP spin columns The high percentage of reads attributed to the host genome in whole metagenome shotgun sequencing experiments makes proper assembly of microbial datasets time consuming and resource intensive, even with sophisticated bioinformatics tools. By removing host DNA in a sample, the QIAamp DNA Microbiome Kit provides samples with enriched bacterial composition for whole metagenome shotgun sequencing. A comparison of reads from whole metagenome shotgun sequencing experiments was performed on DNA isolated from human buccal swabs, where samples were prepared with either the QIAamp DNA Microbiome Kit or solutions from 2 other suppliers. Sequencing of samples prepared with the QIAamp DNA Microbiome Kit resulted in less than 5% human reads, greatly reduced compared to over 90% reads with a kit that does not include host DNA removal and 35% with a kit that does. In a second experiment, samples were spiked with known cultured bacteria. Bacterial DNA were recovered with higher representation compared to a solution without host DNA removal 16S rDNA sequencing is commonly used to determine relative microbial community composition. The QIAamp DNA Microbiome Kit enables the most efficient amplification of the V4 region of DNA purified from buccal swabs when compared to 3 other purification kits (see figure  Efficient amplification and sequencing of 16S rDNA). Sequencing results of the amplified V4 region from 2 samples purified with the QIAamp DNA Microbiome Kit also exhibit the expected bacterial composition representative of a human oral microbiome.

Features

• Fast procedure delivering high-quality total RNA in minutes
• Ready-to-use RNA for high performance in any downstream application
• Consistent RNA yields from small to large amounts of starting material
• No phenol/chloroform extraction, no CsCl gradients, no LiCl or ethanol precipitation
• High-throughput processing in 96-well format and automatable protocols

Product Details

RNeasy Kits are the gold standard for total RNA isolation. They provide fast purification of high-quality RNA from small to large amounts of cells, tissues, and yeast using silica membrane RNeasy spin columns or 96-well plates. Tissue samples can be conveniently stabilized using RNAprotect Tissue Reagent or Allprotect Tissue Reagent, and efficiently disrupted using a TissueRuptor II or TissueLyser II or LT system. The RNeasy 96 Kit enables high-throughput purification of total RNA from up to 96 cultured cell samples using silica membrane RNeasy 96 plates. A dedicated RNeasy QIAcube Kit enables automated purification of 1–12 samples on the QIAcube Connect.

Performance

RNeasy Kits and QIAwave RNA Kits deliver highly reproducible yields of total RNA from small to large samples. Total RNA can reliably be purified from small numbers of cells, including a single cell, as well as from small amounts of standard tissues (see figures  "Reliable RNA isolation from a single cell",  "Highly reproducible yields for sensitive applications" and  "High-quality total RNA from fine needle aspirates") using the RNeasy Micro Kit. RNA purified using the RNeasy Micro Kit provides maximum sensitivity with precious samples in quantitative gene expression analyses, such as real-time RT-PCR, by efficient on-column digestion of genomic DNA (see figure  "Efficient on-column removal of genomic DNA").
Total RNA is easily purified from animal or human cells, animal or human tissues, and yeast (see table “Total RNA yields obtained with RNeasy Kits” and figure  "RT-PCR of RNA from as few as 100 cells").

The QIAwave RNA Mini Kit gives you highly reproducible yields of total RNA from animal or human cells, animal or human tissues, and yeast (see table “Total RNA yields obtained with RNeasy and QIAwave RNA Kits"). The performance between our QIAwave RNA Mini Kit and RNeasy Mini Kit is identical because the chemistry is the same. We’ve also shown that both kits outperform competitors’ kits (see figure   "QIAwave RNA Mini Kit performance").