Actionable insights

NGS Workflow Step 2
Create relevant reports using QIAGEN’s proven gene panels and preeminent bioinformatics platform.
 
Our development team have been hard at work to ensure we can offer you the latest innovations and most relevant content for your GeneReader NGS System. The result? An expanding menu, tailored to address your research needs and deliver only the most actionable cancer insights. Find out how we have optimized our sample prep and bioinformatics solutions to combine liquid biopsy sampling and NGS; Learn how our partnership with Horizon Dx can bring you a standard data set for QC of your entire GeneReader NGS System workflow; Hear what is coming soon

Images
No inhibition in samples derived from anti-coagulated blood.
No inhibition in samples derived from anti-coagulated blood.
[A] and [B] Samples were processed with the MagAttract HMW DNA Kit. Different volumes of eluate were used as indicated. Variables CT values from sample to sample are the results of WBC content variations.
Comparable NGS results using genomic or single-cell amplified DNA
Comparable NGS results obtained using purified genomic DNA or REPLI-g Single Cell amplified DNA.
Whole genome sequencing of the Bacillus subtilis genome was performed on the Illumina MiSeq instrument. For analysis, 2 μg of genomic DNA or DNA amplified from a single cell (three different single cell experiments) and 103 cells, using the REPLI-g Single Cell Kit, was sheared into 300 bp fragments and 1 μg of each was used for library preparation. Comparable sequence coverage was observed for gDNA and REPLI-g Single Cell amplified DNA*. Comparison of nonamplified and REPLI-g amplified DNA revealed error rates in a similar, very low, percentage range.
* Aligned using the Burrows-Wheeler Alignment program (cut-off: 10x coverage): bio-bwa.sourceforge.net.
Comparison on non-amplified and REPLI-g Single Cell amplified DNA also revealed that sequences mapped to the genome with high percentage rates (data not shown
Complete genome coverage.
Complete genome coverage.
Comprehensive analysis of 267 loci spread out over different chromosomes across the entire human genome (as indicated) was performed using RT2 qPCR Primer Assays (QIAGEN) and real-time PCR following DNA amplification with the REPLI-g Single Cell Kit from 3 different single cell experiments. Low and consistent CT values, with no dropout from any marker, indicate that DNA was successfully amplified from all areas of the genome and is highly suited for single-cell genomics.
DNA Purity Depends on Isolation Method
DNA purity depends on the isolation method.
Spectrophotometric scans (220–320 nm) of DNA isolated from leaves/needles using the method of Dellaporta (1), CTAB (2), or the DNeasy Plant Mini Kit. Pure DNA typically shows a symmetrical peak at 260 nm and a smooth profile. Polysaccharides and other secondary metabolites, often copurified with plant DNA isolated using traditional methods, can interfere with OD readings (A260/A280), leading to errors in determination of concentration and purity.
Effective removal of genomic DNA.
Effective removal of genomic DNA.
RNA was isolated from the indicated amounts of RNAlater stabilized rat kidney using the RNeasy Plus Universal Mini Kit or was precipitated using a precipitation reagent from Supplier AII. RNA eluates were analyzed by real-time PCR, with and without reverse transcription, using the QuantiFast Probe RT-PCR Kit and the Rotor-Gene Q.
Efficient long-range PCR
Efficient long-range PCR.
Amplification of a 10 kb fragment of the human ALDH1 gene from genomic DNA isolated from blood. DNA was purified using conventional methods (Phenol) or the QIAamp DNA Blood Maxi Kit (QIAamp). M: 1 kb ladder.
Successful isolation of high-molecular-weight DNA.
Successful isolation of high-molecular-weight DNA.
DNA was isolated from 200 µl of blood stabilized with EDTA, citrate, and heparin respectively, using the standard method recommended for each procedure. The MagAttract HMW DNA Kit outperformed products from other suppliers and enabled successful isolation of 200 kb high-molecular-weight DNA. M: Marker.
Highly efficient rRNA removal
Highly efficient rRNA removal.
[A] The lack of any rRNA peak in QIAxcel trace data demonstrates complete removal of rRNA in the rRNA-depleted sample, in contrast to the total RNA sample. [B] qRT-PCR assays indicate that more than 99% of all rRNA species, including mitochondrial rRNA, are removed with the kit.
High-quality RNA
High-quality RNA.
RNA was purified from rat tissue using the RNeasy Plus Universal Mini Kit with or without gDNA Eliminator Solution. [A] Real-time RT-PCR analysis was carried out using RNA isolated from 25 mg frozen liver tissue. The CT value of all samples was comparable. DNA contamination was detected in the -RT control if gDNA Eliminator Solution was not used, whereas the samples isolated using gDNA Eliminator Solution showed minor or no DNA contamination. [B] RNA was purified from 25 mg stabilized heart tissues. Data obtained using the QIAxcel system demonstrates good RNA integrity regardless of whether RNA was isolated using gDNA Eliminator Solution.
Innovative UV treatment eliminates any detectable trace of residual DNA in kit components
Innovative UV treatment.
Bacterial DNA (2000 copies) was spiked into REPLI-g sc Reaction Buffer, which was then irradiated with UV using the standard procedure for all buffers and reagents provided with the REPLI-g Single Cell Kit (following UV treatment, the kits undergo stringent quality control to ensure complete functionality). In subsequent real-time PCR, no bacterial DNA was detectable following the UV decontamination procedure.
Next-generation sequencing using REPLI-g amplified DNA requires less hands-on time and generates more sequence information than PCR-based methods
Less hands-on time and more sequence information.
PCR-based whole genome amplification (WGA) and library preparation for next-generation sequencing requires a purification step prior to library preparation that can result in ~3 times more hands-on time than REPLI-g based WGA and library preparation. Additionally, unlike REPLI-g amplified DNA, PCR-based methods also include PCR primer binding sites (indicated in red) on the WGA amplification product. Since next-generation sequencing read-lengths are between 50–200 bp, the resulting genome coverage could be strongly reduced using a PCR-based WGA method.
Multiple Displacement Amplification (MDA) technology delivers long read lengths with isothermal amplification.
Multiple Displacement Amplification (MDA) technology delivers long read lengths with isothermal amplification.
Primers (arrows) anneal to the template DNA and are extended at 30°C by Phi 29 polymerase, which moves along the DNA template strand, displacing the complementary strand while becoming a template itself for replication. In contrast to PCR amplification, MDA does not require different temperatures and ends in very long fragments with low mutation rates.
Purified Genomic DNA up to 50 kb  Size distribution of DNA prepared with QIAamp Kits from the indicated sources (3 µg per lane)
Purified genomic DNA up to 50 kb.
Size distribution of DNA prepared with QIAamp Kits from the indicated sources (3 µg per lane).
Automate your NGS purification.
The innovative QIAcube uses advanced technology to process QIAGEN spin columns, enabling seamless integration of automated sample preparation into your NGS workflow.
The REPLI-g Single Cell Kit delivers unbiased amplification of DNA from a single cell
The REPLI-g Single Cell Kit delivers highly reliable, unbiased amplification of DNA from a single cell.
The REPLI-g Single Cell Kit or single cell kits from Suppliers G, N, and S were used to individually amplify 5 human cells. Real-time PCR was used to analyze 3 markers to identify loss or variability in the amount of genomic loci. Unlike kits from other suppliers, the REPLI-g Single Cell Kit delivered unbiased amplification of DNA in each of the 5 cells, indicated by equivalent CT values for each marker. Unlike with the REPLI-g Single Cell Kit, DNA amplified using the kits from Suppliers G and N demonstrated high dropout rates. For both kits, genomic marker X54 was not amplified in 2 of the 5 cells tested, and the kit from Supplier G did not amplify marker 99 in 1 of the 5 cells, indicating incomplete genome coverage and biased amplification that makes these kits unsuitable for reliable single cell research.