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GALENVS MagnetiQ Operating instructions

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BC0100-10 / BC0250-10 / BC/0096-10BC0016-10 v1.0
Blood & Cell DNA Extraction kit
Introduction
The Galenvs magnetiQ Blood & Cell DNA Extraction Kit enables a rapid and efficient method of genomic
DNA (gDNA) isolation and purification from whole blood samples and mammalian cell suspensions. Our
magnetic bead technology allows for a combined lysis and capture step and efficient washing steps leading
to high yields of concentrated gDNA free of cell debris, salts, lipid, or protein contamination under 20
minutes.
Applications
• Next Generation Sequencing (NGS)
• PCR and qPCR gene amplification
• DNA methylation studies
• Copy number variation (CNV) studies
Features
• Amenable to high-throughput methods and automation (Kingfisher Flex, Allsheng
AutoPure96, etc.)
• Faster and simpler magnetic collection and resuspension steps
• Reduced clogging and contaminant/cell debris carryover
• Eliminates organic solvent hazardous waste
• Requires only the use of a magnetic rack and standard pipettes
• Does not require centrifugation steps
• Storage and operation at room temperature
Performance
Yield and Purity
In comparing the Galenvs magnetiQ Blood & Cell kit to industry leading products (DNeasy Blood & Tissue
by Qiagen), 100 L of whole human blood (Potassium/EDTA stabilized) are processed as described by
manufacturer recommended protocols (Figure 1).
BC0100-10 / BC0250-10 / BC/0096-10BC0016-10 v1.0
Figure 1: Extraction yield of gDNA from 100 L of whole blood using Galenvs magnetiQ Blood/Cell kit and
Qiagen DNeasy Blood & Tissue kit. Quantification of gDNA performed using Qubit BR DNA fluorescent assay
measurement (n=3)
Table 1 : A260/A280 purity ratios of eluted gDNA samples from 100 L whole blood measured by UV/Vis nanodrop
(n=3)
A260/A280
Galenvs
1.83 0.03
Qiagen
2.04 0.12
0
5
10
15
20
25
30
35
40
45
50
Galenvs Qiagen
µg/µL
gDNA Yield
BC0100-10 / BC0250-10 / BC/0096-10BC0016-10 v1.0
-100
400
900
1400
1900
2400
15 20 25 30 35 40
RFU
Cycle
pre-GAPDH Quantification
Galenvs
Qiagen
qPCR
Both quantitative PCR (qPCR) (Figure 2) and gel electrophoresis results (Figure 3) confirm the higher recovery yields
obtained with Qubit BR DNA assay. Eluted gDNA is added to the qPCR master mix (NEB Luna) containing primers
for pre-GAPDH housekeeping gene.
Figure 2: Amplification of the pre-GAPDH gene by qPCR from 100 L of whole blood comparing Galenvs and
Qiagen extraction kits
Figure 3: Gel electrophoresis of extracted gDNA from 100 L whole blood samples (4x) comparing Galenvs and
Qiagen kits. Galenvs isolated gDNA is higher molecular weight while Qiagen extraction shows DNA smears below
gDNA bands
CT
Galenvs
23.52
Qiagen
24.48
BC0100-10 / BC0250-10 / BC/0096-10BC0016-10 v1.0
Sample Volume
To illustrate the scalability of the Galenvs protocol, higher volumes of whole blood were processed while scaling the
lysis buffer volume accordingly. The elution volumes can be further tuned to obtain the desired final concentration
of isolated gDNA. Figures 4 and 5 show the yield and purity, respectively, from varying volumes of whole blood. The
total yield of gDNA increases linearly and matches the scaling factor of processed whole blood volume – while
maintaining high purity ratios. This is also confirmed in the gel electrophoresis analysis (Figure 6).
Figure 4: Total extraction yield of gDNA from varying
whole blood volumes
Figure 5: Extracted gDNA purity from whole blood
samples of different volumes
Figure 6: Gel electrophoresis of extracted gDNA from 100 L, 300 L, and 1000 L of whole blood samples
processed with Galenvs magnetiQ kit
Specifications
1.85 1.89 1.91
0
0.5
1
1.5
2
2.5
100 µL 300 µL 1000 µL
Blood Sample Volume (µL)
A260/A280
BC0100-10 / BC0250-10 / BC/0096-10BC0016-10 v1.0
Sample Type
Blood, Cells, Bacteria
Quantity
100 or 250 assays / Prefilled plates (16-96 preps)
Elution Volume
100 µl
Processing Mode
Automated – Manual
Sample Volume
100 µL – 1000 µL
Binding Technology
Magnetic beads
Binding Capacity
Scalable
Components
(i) Lysis/Binding Buffer
(ii) Wash Buffer #1
(iii) Wash Buffer #2
(iv) Elution Buffer
(v) Proteinase K
Storage
Room temperature
Product codes
100 preps
BC0100
250 Preps
BC0250
Prefilled plates 96-well
BC0096
Prefilled plates 16-well
BC0016
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