Sample Technologies

Innovative technologies deliver high yields of high-quality nucleic acids

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High quality starting material is essential for reliable next-generation sequencing results and accurate data analysis. QIAGEN, with leading expertise in sample preparation technologies, offers solutions that deliver high yields of superior-quality nucleic acids and enable outstanding results — on any sequencing platform.

QIAGEN's wide range of sample technologies delivers:

High-quality DNA and RNA purification, even from challenging sample types
Unbiased whole genome amplification from a single cell
Time savings and standardization with automated purification

DNAseq NGS workflow
DNA purification from challenging samples
Unbiased whole genome amplification from single cells and limited sample material

RNAseq NGS workflow
High-quality RNA purification

Automation for all NGS workflows
Automated sample purification on the QIAcube

Sample purification is a critical step for obtaining accurate NGS data (see figure DNA purity depends on the isolation method). Even the best and most robust NGS technologies can only provide reliable results if the template nucleic acid is of adequate quality and purity. QIAGEN’s diverse portfolio of sample prep solutions is especially adapted to overcome all challenges in DNA and RNA purification, including low yields, poor quality, and difficult sample types, such as formalin-fixed, paraffin-embedded (FFPE) samples (see table "Wide range of purification kits). All QIAGEN NGS sample prep solutions can be easily automated on the QIAcube, ensuring greater standardization and time savings.

Wide range of purification kits
Sample type	Nucleic acid	Kit
Plant cells or tissues	DNA	DNeasy Plant Mini Kit
Blood	DNA	QIAamp DNA Blood Mini Kit
Tissues, swabs, CSF, blood, body fluids	DNA	QIAamp DNA Mini Kit
High-molecular-weight DNA	DNA	MagAttract HMW DNA Kit
Cells, easy-to-lyse tissues, plant	RNA	RNeasy Plus Kits
All types of difficult to lyse tissues	RNA	RNeasy Plus Universal Kits
FFPE tissue	RNA	RNeasy FFPE Kit

DNAseq NGS workflow
High-quality DNA purification — even from challenging sample types
For use with all sample types, QIAamp and DNeasy Kits deliver consistent, high yields of DNA with complete removal of contaminants and inhibitors (see table "Yields with the QIAamp DNA Mini Kit"). Purified DNA is up to 50 kb, making it highly suited for long-range PCR amplification and NGS applications (see figure Efficient long-range PCR and Purified genomic DNA up to 50 kb).

Yields with the QIAamp DNA Mini Kit
		Yield
Sample	Starting amount	Total nucleic acids (µg)^†	DNA (µg)^‡
Blood	(200 µl)	4–12	4–12
Buffy coat	(200 µl)	25–50	25–50
Cells	(10⁷)	40–60	30–40
Liver	(25 mg)	60–115	10–30
Brain	(25 mg)	35–60	15–30
Lung	(25 mg)	25–45	5–10
Heart	(25 mg)	15–40	5–10
Kidney	(25 mg)	40–85	15–30
Spleen	(10 mg)	25–45	5–30

Purification of high-molecular-weight DNA
For sensitive applications like NGS, it is essential that inhibitors, such as anticoagulants, enzymes, and divalent cations, are completely eliminated from purified DNA. The MagAttract HMW DNA Kit successfully isolates high-molecular-weight (HMW) DNA (100–200 kb) using a simple, magnetic bead-based protocol, and delivers high yields of DNA that is especially suited for NGS applications (see figure Successful isolation of high-molecular-weight DNA). Real-time PCR amplification of DNA using the MagAttract HMW DNA Kit demonstrates the complete lack of inhibition in all samples (see figure No inhibition in samples derived from anti-coagulated blood).

Unbiased whole genome amplification from single cells and limited sample material, recommended for use in NGS
The REPLI-g Single Cell Kit uniformly amplifies gDNA from single cells without purification (<1000 cells to as little as 1 bacterial or tumor cell) or purified gDNA. Negligible sequence bias and maximized genome coverage make the REPLI-g Single Cell Kit an excellent tool for a wide range of NGS applications, including cancer research, metagenomics, and stem cell biology.

DNA sequence analysis of biological samples using innovative instrumentation, such as next-generation sequencing platforms, is often limited by the small amount of sample available. DNA amplified using the REPLI-g Single Cell Kit has been tested with, and is highly suited for, numerous downstream analyses, including NGS. Since there is no requirement for a separate PCR-based amplification step, REPLI-g whole genome amplification and library preparation requires less hands-on time and results in longer read-lengths than PCR-based methods (see figure Less hands-on time and more sequence information). High-quality, comparable NGS results showing a high percentage of sequence coverage and very low error rates are achieved with both purified genomic DNA or REPLI-g Single Cell amplified DNA, including when starting from just a single bacterial cell (see figure Comparable NGS results). These findings are underscored by a comprehensive analysis of a wide range of markers covering all human autosomal chromosomes and the X chromosome, with 3 different independent experiments demonstrating that DNA is successfully amplified from all areas of the genome without a single drop-out (see figure Complete genome coverage). Unlike with the REPLI-g Single Cell Kit, single cells analyzed using kits from other suppliers often fail in complete and unbiased sequence representation (see figure Unbiased DNA amplification from a single cell).

Unbiased amplification from a single cell is achieved with Multiple Displacement Amplification (MDA) technology and a modified form of Phi 29 Polymerase (see figure Multiple Displacement Amplification (MDA) technology, along with a unique, controlled decontamination procedure to avoid amplification of contaminating DNA, ensuring highly reliable results each time (see figure Innovative UV treatment). For more detailed information on REPLI-g whole genome amplification, see our WGA spotlight page.

RNAseq NGS workflow
High-quality RNA purification
Purification of high-quality RNA can be challenging due to rapid degradation, and is an important consideration in your RNAseq workflow. RNeasy is the standard technology for high-quality RNA purification. RNeasy Plus Kits integrate fast, convenient RNA purification with effective elimination of genomic DNA (gDNA), while RNeasy Plus Universal Kits are especially suited for difficult-to-lyse tissues. High, reproducible RNA yields are achieved from a wide range of sample materials (see table "Typical yields of total RNA with RNeasy Plus Universal Kits").

Typical yields of total RNA with RNeasy Plus Universal Kits
Mouse/rat tissue (10 mg)	Yield of total RNA (µg)*
Adipose tissue	0.5–2.5
Brain	5–20
Heart	5–25
Intestine	10–60
Kidney	5–40
Liver	15–80
Lung	5–15
Muscle	5–35
Skin	2–5
Spleen	15–100

Analysis of purified RNA using the QIAxcel system demonstrates excellent RNA integrity (see figure High-quality RNA). gDNA Eliminator Solution and RNeasy technologies efficiently remove most gDNA contamination without DNase treatment (see figure Effective removal of genomic DNA).

RNeasy Plus Kits, in combination with the GeneRead rRNA Depletion Kit, deliver highly pure RNA that is especially suited for NGS applications — with more than 99.5% of rRNA depleted from the sample (see figure Highly efficient rRNA removal).

Automation for all NGS workflows
Save time and standardize your NGS results with automated sample purification on the QIAcube
The innovative QIAcube uses advanced technology to process QIAGEN spin columns, enabling seamless integration of automated, low-throughput sample prep into your laboratory workflow. No change of purification chemistry is required, assuring fast startup and immediate results. All steps in the purification procedure are fully automated — and up to 12 samples can be processed per run. The QIAcube fully automates more than 40 QIAGEN spin-column kits using a simple lyse, bind, wash, elute procedure and is an important component of the GeneReader Sample to Insight workflow.