High-throughput - Omega Bio-tek

HIGH THROUGHPUT NUCLEIC
ACID PURIFICATION SYSTEMS
nucleic acid purification utilizing magnetic beads
& silica plate technologies
Innovations in Nucleic Acid Isolation
COMPANY PROFILE
Since its founding in 1998, Omega Bio-tek, Inc. has been at the forefront of nucleic acid purification by offering
products for clinical and basic research, biotechnology and agricultural applications. DNA and RNA extraction is the first
step for so many downstream analyses, and efficient and clean nucleic acid isolation is crucial. Our goal is to offer high
quality products to help you improve your workflows. Our diverse product line ranges from RNA purification from
plants to DNA extraction from dried blood spots. By
offering over 900 products for nucleic acid isolation,
we have a solution to almost any of your nucleic acid
purification needs. Contact one of our specialists to
see which product would best fit your application.
With the ability to offer individual components and
customize kits, we can help your lab reduce waste and
increase productivity.
Quality is key to operations at Omega Bio-tek. We
offer products for manual and automated
processing and we support our customers by having
Hamilton STAR®, Beckman Coulter Biomek® FX and
Thermo KingFisher® instruments in house. In addition
to offering nucleic acid purification products, we also
offer next generation sequencing services utilizing
Illumina platforms. We are ISO 9001:2008 certified and
we ensure that our products are properly assembled, tested, recorded, stored and shipped. We perform rigorous quality
checks of our products and thoroughly train our employees to ensure compliance. We also have several quality control
steps within our processes to deliver the best product. We firmly believe that quality in equals quality out.
THE OMEGA DIFFERENCE
•
ECONOMICAL:
On average, Omega Bio-tek’s
products cost 30% less than the competition.
•
CUSTOMIZABLE:
For large customers, we can generate custom
packaging, kits, reagents, etc.
•
INDIVIDUAL COMPONENTS:
We sell kit components separately.
•
PRODUCT RANGE:
Alternative options for almost all of your nucleic acid
purification needs.
01
HIGH THROUGHPUT PRODUCTS
MAG-BIND® PARTICLES
Our Mag-Bind® technology provides a flexible and
reliable method for isolating nucleic acids using
magnetic beads. Our magnetic bead-based kits are
optimized in processing times and binding capacity for
each application. Magnetic beads are available in 24-,
96- or 384-well formats to fit your sample size. The same
kit can be modified for different formats by changing
the volumes, plasticware and magnetic stands.
Magnetic beads allow for higher recovery and
yield of nucleic acids versus filter-based methods.
Magnetic stands with SBS standard format
can be added to liquid handlers, which can be
specifically programmed for an inexpensive
way to process multiple plates simultaneously.
Mag-Bind® technology is ideally suited for
liquid handlers that have a 96-channel head, but it
can be adapted to 8-channel head instruments.
Mag-Bind® kits can also be used with Thermo
KingFisher Flex®, Thermo KingFisher DUO®, Qiagen
BioSprint® and Life Technologies MagMAX® 96.
MAG-BIND® PARTICLES
SILICA PLATE TECHNOLOGY
Omega Bio-tek HiBind® technology is the basis for our E-Z 96® silica plates that have a binding capacity of up
to 50 µg of genomic DNA or total RNA and 20 µg of plasmid DNA. Our kits use semi-skirted 96-well plates that
fit manual and automated vacuum manifolds and plate adaptors for centrifugation.
The E-Z 96® silica plates have a silica glass fiber matrix that offers excellent well-to-well consistency and
includes tip directors to eliminate cross contamination. The plates feature a 750 µL well capacity to reduce
sample processing time. The E-Z 96® silica plates are an ideal option for liquid handlers with 8- or 96-channel
pipettors.
VACUUM MANIFOLDS
The E-Z 96 vacuum manifold is designed for manual
DNA/RNA sample preparation with E-Z 96® DNA/RNA
plates. The E-Z vacuum manifold is also compatible
with other semi-skirted filter plates.
The upper plate has a molded recess that fits our
filter plate and contains a custom gasket to create
a secure, airtight connection to the filter plate. The
plates have pins and holes for easy alignment and
stability.
02
LIBRARY NORMALIZATION
MAG-BIND® EQUIPURE LIBRARY &
gDNA NORMALIZATION KITS
Many high throughput applications, such as sequencing and genotyping, require the input DNA
concentration to be within a certain range for optimal results. Traditionally, a tedious process of quantification,
calculation and concentration adjustment must be carried out to normalize the DNA samples.
The Mag-Bind® EquiPure Normalization Kit completely eliminates the need to quantify and aliquot DNA, saving time, quantification bias and tip cost. Using our proprietary Mag-Bind® Normalizer Beads and binding buffer system, input DNA of various quantities is simply bound, washed and eluted to a final normalized product.
The magnetic beads have a limited binding capacity and therefore allow a pre-defined amount of DNA to be
captured and eluted. An alternate protocol allows for non-bound DNA to be recovered.
The Mag-Bind® EquiPure Normalization Kit allows users to adjust the quantity of DNA that binds to the magneticc beads and elution volumes can be adjusted to vary DNA concentrations to user’s needs.
The Mag-Bind® EquiPure Normalization Kit is fully automatable on multiple liquid handling platforms including Hamilton STAR, Tecan Evo, Caliper Sciclone and Beckman Coulter Biomek instruments.
NORMALIZATION OF 16S
NGS LIBRARIES
NORMALIZATION OF RNA-SEQ
NGS LIBRARIES
900
800
150
1500
700
2000
250
120
2000
DNA (ng)
DNA (ng)
600
500
400
90
60
300
30
200
100
0
0
Input DNA Samples
Varying amounts of 16s NGS libraries were prepared and
normalized with the Mag-Bind® EquiPure Normalization
Kit. DNA yield was quantified using Promega QuantiFluor®
dsDNA system.
03
Input DNA Samples
Varying amounts of Illumina TruSeq Stranded mRNA
libraries were prepared and normalized with the
Mag-Bind® EquiPure Normalization Kit. DNA yield
was quantified using Promega QuantiFluor® dsDNA
system.
gDNA NORMALIZATION
qPCR FROM NORMALIZED gDNA
100
90
80
70
60
50
40
30
20
10
0
30
29
28
27
cT
DNA Output (ng)
NORMALIZED gDNA
26
25
24
23
22
21
500
750
1,000
2,000
20
500
750
1,000
2,000
DNA Input (ng)
Genomic DNA was isolated from whole blood with the Mag-Bind® Blood & Tissue DNA HDQ Kit. Varying amounts were inputted into the
different reactions then normalized with Mag-Bind® EquiPure gDNA Kit. DNA yield was quantified using Promega QuantiFluor® dsDNA
system.
Mag-Bind® EquiPure Library
Normalization Kit
Product
M6445-00
M6445-01
Preparations
1 x 96
4 x 96
Mag-Bind® EquiPure gDNA
Normalization Kit
Product
M6423-00
M6423-01
Preparations
1 x 96
4 x 96
04
DYE TERMINATOR REMOVAL
MAG-BIND® SEQDTR
Omega Bio-tek’s Mag-Bind® SeqDTR is designed to effectively and reliably remove unincorporated
terminators from sequencing reactions. Sequencing products are mixed with the Mag-Bind® SeqDTR
magnetic particles which selectively bind DNA. Two rapid wash steps eliminate trace contaminants such
as nucleotides and primers to reduce background signal and therefore achieve higher QV scores. The high
sensitivity of Mag-Bind® SeqDTR’s binding ability allows for decreased concentrations of BigDye® chemistry to
be used and longer continuous read lengths to be achieved. Mag-Bind® SeqDTR can be processed in 96- and
384-well formats and is compatible with many liquid handling instruments including Hamilton STAR/STARlet,
Beckman Coulter Biomek® FX/NX, and Tecan Evo® instruments. Up to four plates can be run in a 96-well format
in less than 25 minutes.
Mag Bind SEQ DTR
Company C
Company A
SIGNAL STRENGTH
1200
900
1000
850
800
800
600
750
400
700
200
650
0
600
CONTINUOUS READ
LENGTH
60
QUALITY VALUE
SCORE
55
50
45
40
Comparison of magnetic bead-based big dye clean-up chemistries. Purified 1.8 kb PCR fragments were sequenced from each
company using the recommended protocols. The median of 16 samples per company are used in the representations above. A 5 µL
sequencing reaction was performed using a 1/32 dilution of Applied Biosystems Big Dye Terminator v3.1 Chemistry. DNA was analyzed
on an Applied Biosystems 3730XL.
Mag-Bind® SeqDTR
Product
05
Size
M1300-05
5 mL
M1300-08
50 mL
M1300-50
500 mL
PCR PURIFICATION
MAG-BIND® RXNPURE PLUS
Omega Bio-tek’s Mag-Bind® RxnPure Plus Kit allows for the
rapid and reliable recovery of DNA from PCR and other
enzymatic reactions. The system combines Omega
Bio-tek’s proprietary chemistries with the reversible
nucleic acid binding properties of magnetic beads to
selectively bind PCR amplicons 100 bp and larger to
eliminate excess nucleotides, primers and small, nontargeted amplification products, such as primer dimers.
This kit is designed for manual and fully automated
purification and may not require programming of current
liquid handling instruments depending on your current
method.
Mag-Bind® RxnPure Plus is compatible with multiple
liquid handlers and magnetic processors such as Hamilton
STAR®/STARlet® and Beckman Coulter Biomek® FX/NX.
RxnPure
Plus
Company
A
50 BP
Ladder
Mag-Bind® RxnPure Plus
Ten microliters of 50 bp ladder was purified with
Omega Bio-tek’s Mag-Bind® RxnPure Plus Kit and
a comparable kit from Company A, according to
manufacturer’s recommended protocols. The DNA
was eluted in 20 microliters and analyzed on the
Agilent Tapestation 2200.
Product
Size
M1386-00
5 mL
M1386-01
50 mL
M1386-02
500 mL
06
PLASMID DNA
E-Z 96® FASTFILTER PLASMID DNA KIT
The E-Z 96® FastFilter Plasmid DNA Kit combines the
power of HiBind® technology with the time-tested
consistency of alkaline-SDS lysis of bacterial cells to
deliver high quality plasmid DNA. Using the E-Z 96® DNA
plate, up to 96 samples can be processed in less than 90
minutes. The multi-layered E-Z 96® Lysate Clearance Plate
is designed to eliminate time-consuming centrifugation
steps for the clearance of bacterial alkaline lysates without
clogging. With the combination of these two plates, this
kit boasts an average DNA recovery rate of 10-30% higher
than traditional manual centrifugation methods.
The E-Z 96® FastFilter Plasmid DNA Kit typically yields
10-15 µg high-copy plasmid DNA from a 1 mL overnight
culture, although yields will vary according to plasmid
copy number, E. coli strain and growth conditions. The 96well format of the E-Z 96® FastFilter Plasmid DNA Kit lends
itself to automation on platforms equipped with vacuum
manifolds such as Hamilton Nimbus®, STAR® and STARlet®
lines, Beckman Coulter Biomek® FX and NX workstations
and Tecan Freedom Evo® systems.
E-Z 96 FastFilter Plasmid DNA Kit
Product
Preparations
D1097-00
1 x 96
D1097-01
4 x 96
D1097-02
20 x 96
07
AUTOMATED EXTRACTION ON
HAMILTON STAR
MANUAL EXTRACTION
Extraction
Method
Automated
Manual
Conc.
(ng/µg)
77.1
81.7
A260/A280 A260/A230
1.89
1.91
1.80
1.81
Comparison of automated and manual extraction methods using
the E-Z 96 FastFilter Plasmid DNA Kit. Ninety-six 1.2 mL overnight
cultures in 2xYT medium were lysed in a modified alkaline lysis system and
cleared with an Omega Bio-tek E-Z 96 Lysate Clearance Plate. DNA was
extracted from one half of the samples using the standard manual
method and the other half using the Hamilton STAR liquid handling
workstation.The samples were run on a 1% agarose gel. Concentration and
quality was quantified using Thermo Fisher NanoDrop® 2000c. The table
represents the average of 48 samples.
PLASMID DNA
MAG-BIND® ULTRA PURE PLASMID DNA 96 KIT
The Mag-Bind® Ultra Pure Plasmid DNA 96 Kit combines our alkaline lysis method with magnetic beads to offer a fast
and economical option for high-quality plasmid DNA extraction. Two options are available to clear cellular debris after
alkaline lysis: centrifugation or lysate clearing beads. The lysate clearing magnetic beads allow for fully automated
processing after pelleting of the cells from culture. The kit utilizes Omega Bio-tek’s endo-free plasmid DNA binding
system for purification of high-quality DNA that is endotoxin-free.
This flexible system can easily be adapted to most automated liquid handling platforms without sacrificing plasmid
DNA quality. Plasmid DNA is suitable for restriction enzyme digestion, Sanger sequencing and transfection.
ENDOTOXINS IN PLASMID
DNA PREPS
PLASMID DNA YIELD COMPARISON
15
Prep Method
EU/µg
Mag-Bind® Ultra-Pure Plasmid DNA Kit
Company P Magnetic Bead Plasmid DNA Kit
Omega Bio-tek E.Z.N.A.® Plasmid Mini Kit 1
0.05
11.79
12.94
12
9
6
Endotoxins in plasmid DNA preps. Plasmid DNA was
isolated from 0.8 mL LB culture from the corresponding
method following the manufacturer’s recommended protocols.
Endotoxin level was determined with Thermo Scientific’s Pierce
LAL Chromogenic Endotoxin Quantitation Kit.
3
0
Omega Bio-tek E.Z.N.A.® Plasmid DNA Kit 1
Omega Bio-tek Mag-Bind® Ultra Pure Plasmid DNA 96
Kit with lysate clearance via centrifugation
Omega Bio-tek Mag-Bind® Ultra Pure Plasmid DNA 96
Kit with lysate clearance via lysate clearing beads
Company Q Silica Spin Columns
Company P Magnetic Bead Plasmid DNA Kit
Mag-Bind® Plasmid DNA 96 Kit
Product
M1258-00
M1258-02
Preparations
1 x 96
4 x 96
Plasmid DNA yield comparison. Plasmid DNA was
isolated according to manufacturer’s recommended
protocols from 0.8 mL LB cultures grown for
24 hours.
Each protocol was performed in
triplicate with the average data shown above.
DNA was quantified with Thermo Scientific
NanoDrop 2000c.
08
BLOOD, TISSUE, SALIVA & BUCCAL SWABS DNA
MAG-BIND® BLOOD & TISSUE DNA HDQ 96 KIT
The Mag-Bind® Blood and Tissue DNA HDQ 96 Kit is designed for the rapid and reliable isolation of highquality genomic DNA from 10-250 µL of blood, buccal swabs, buffy coat, or cultured cell samples. The
magnetic response time of the Mag-Bind® particles and the elimination of all heating steps allow for an overall
rapid processing time.
Using paramagnetic particles provides high-quality DNA that is suitable for direct use in most downstream
applications such as qPCR, PCR, and next generation sequencing.
The Mag-Bind® Blood and Tissue DNA HDQ 96 Kit is compatible with multiple liquid handlers and
magnetic processors such asThermo KingFisher Flex, Qiagen BioSprint, Applied Biosystems/Ambion MagMAX 96,
Beckman Coulter Biomek FX/NX, and Hamilton Star/Starlet.
WHOLE BLOOD DNA
ISOLATION FROM 1,500 µL
1
2
3
4
5
6
Nucleic
Acid
Conc.
118.4
90.8
80.5
127.2
73.1
1.83
A260/A280
A260/A230
Yield
29
1.82
1.84
1.85
1.81
1.75
1.83
2.09
2.9
2.83
2.19
1.18
2.48
59.2
45.4
40.3
63.6
36.5
81.4
25
27
cT
Sample
qPCR FROM gDNA ISOLATED
FROM WHOLE BLOOD
23
21
19
17
15
50 ng
5 ng
0.5 ng
Template Amount
Genomic DNA was extracted from 1,500 µL whole blood using Omega Bio-tek’s Mag-Bind® Blood and Tissue DNA HDQ 96 Kit. DNA was analyzed
on a Thermo Scientific NanoDrop® 2000c. Fifty nanograms of DNA was diluted 10 and 100-fold and used as a template in a 20 microliter SYBR qPCR
reaction. The cT values increased by only three cycles per 10-fold dilution, which demonstrates that the template DNA is free of inhibition.
Mag-Bind® Blood & Tissue DNA HDQ 96 Kit
Product
M6399-00
M6399-01
09
Preparations
1 x 96
4 x 96
BLOOD, TISSUE, SALIVA & BUCCAL SWABS DNA
10
DNA YIELD FROM 200 µL
WHOLE BLOOD
Yield (µg)
8
6
4
SALIVA DNA EXTRACTION
Kit
Biomatrica
Oragene
Omega Bio-tek
48
55
Company A
43.5
28
Saliva DNA extraction. DNA yield from 200 µL saliva
samples stabilized in Biomatrica’s DNAguard and DNA Genotek
Oragene saliva tubes. DNA was extracted in quadriplicate
according to the manufacturer’s recommended protocols
and eluted in equal amounts. DNA yield was determined by
PicoGreen quantification and average DNA concentration is
shown.
2
0
Samples
DNA yield from 200 µL whole blood. DNA yield from 200 µL
whole blood extracted using the Mag-Bind® DNA HDQ Kit on
the ABI MagMAX 96 instrument. DNA yield was determined by
PicoGreen quantification.
BUFFY COAT DNA EXTRACTION
Sample
Yield
UV
Yield
flourescent dye
Omega Bio-tek
34.5
41
Omega Bio-tek
35.1
41
Omega Bio-tek
42.9
48
Company T
45.3
14
Company T
41.9
19
Company T
40.3
13
Buffy coat DNA extraction.
1,000 µL buffy coat
samples’ DNA was extracted in triplicate according to the
manufacturer’s recommended protocols and eluted in equal
amounts. DNAyieldwasdeterminedbyPromegaQuantusdsDNA
quantification kit (flourescent dye) and Thermo Scientific
Nanodrop 2000c (UV).
BUCCAL SWABS DNA
EXTRACTION COMPARISON
Omega Bio-tek
Company A
Genomic DNA was extracted from buccal swab samples
using Omega’s Mag-Bind® Blood DNA HDQ 96 Kit and
Company A’s genomic DNA extraction kit. Ten percent
of DNA extracted from each sample was analyzed on a
0.8% agarose gel.
10
VIRAL DNA & RNA
MAG-BIND® VIRAL DNA/RNA 96 KIT
The Mag-Bind® Viral DNA/RNA 96 Kit isolates DNA and RNA from serum, plasma, swabs, cell culture
supernatants, urine and other acellular body fluids. This kit uses a unique system to efficiently lyse viral
samples. The high binding ability and sensitivity of the magnetic particle technology allows efficient isolation
from low titers or large sample sizes.
The samples are lysed in a specially formulated buffer containing detergent. Binding conditions are adjusted
to enable the nucleic acid to bind to the Mag-Bind® particles. After proteins and cellular debris are efficiently
washed away, pure RNA and DNA is eluted in nuclease-free water or Elution Buffer. This system can be used
on multiple automated platforms including Ambion MagMAX™, Thermo KingFisher® Flex, Beckman Coulter
Biomek® FX, and Hamilton STAR instruments.
RNA Template
40
1 x 10 viral particles/µL
7
35
30
cT
25
1 x 106 viral particles/µL
20
15
10
1 x 105 viral particles/µL
5
0
Omega Bio-tek
Company A
1 x 104 viral particles/µL
HBV virus (in quantities of 10 and 1 infectious unit(s)) was
spiked into 200 µl of human serum. Viral nucleic acid was
isolated with Omega Bio-tek’s Mag-Bind® kit and with a
comparable kit from Company A according to
recommended protocols. 5 µl of template was used for a
SYBR Green labeled qPCR reaction, which was replicated
four times. The resulting mean cT values are shown in the
above figure.
Mag-Bind® Viral RNA/DNA 96 Kit
Product
Preparations
M6246-01
1 x 96
M6246-02
4 x 96
M6246-03
12 x 96
11
1 x 103 viral particles/µL
cT Value
19.90
19.88
19.98
23.09
23.09
22.99
25.48
25.53
25.08
28.64
28.56
28.66
31.23
31.59
31.58
cT values of recovered RNA isolated using the Mag-Bind®
Viral RNA/DNA 96 Kit. Serum was separated from a human
blood sample containing 1 x 107 Hepatitis B viral particles/
µL. A 10-fold dilution series of the serum was performed and
50 µL of each dilution was used in the Mag-Bind® Viral RNA/
DNA 96 Kit to isolate viral RNA. Two µL RNA was used as the
template.
PATHOGEN DNA & RNA
MAG-BIND® UNIVERSAL PATHOGEN KIT
The Mag-Bind® Universal Pathogen 96 Kit allows for
the rapid and reliable isolation of high quality host
genomic DNA, Gram-positive and negative bacterial
DNA, fungal spore DNA, viral DNA and viral RNA from
tissue, blood, urine, serum and fecal samples.
E-Z 96® Disruptor Plus plates are included with the
kit for simultaneous homogenization and lysis of
the sample. The unique buffer system does not
require alcohol to bind nucleic acids, allowing
recovery of high quality DNA/RNA free of PCR
inhibitors. The entire procedure can be completed in
less than 60 minutes for 96 samples.
Undiluted
1:10
37
36
Average cT
The system allows for automation after sample lysis
via Hamilton STAR, Thermo KingFisher Flex, Applied
Biosystems MagMAX, Qiagen BioSprint and other
liquid handling instruments.
GRAM-POSITIVE BACTERIA
35
34
33
32
31
Stool
Urine
Group B strep cultured samples were added to
corresponding sample types and isolated with MagBind® Universal Pathogen Kit. 20 µL SYBR qPCR were
performed in triplicate on primers specific to the target
organism. Average of triplicate data shown.
FUNGAL SPORES
Undiluted
1:10
1:100
30
VIRAL RNA
20
1:10
30
10
25
5
0
Undiluted
35
15
Serum + Crypto Stool + Crypto Urine + Crypto
Cryptosporidium oocysts were added to corresponding sample
types and isolated with Mag-Bind Universal Pathogen Kit. 20 µL
SYBR qPCR were performed in triplicate on primers specific for
the target organism. Average of triplicate data shown.
Average cT
Average cT
25
20
15
10
5
0
Mag-Bind® Universal Pathogen 96 Kit
Product
M4029-00
M4029-01
Preparations
1 x 96
4 x 96
Stool
Urine
Serum
HBV viruses were added to corresponding sample types and
isolated with Mag-Bind® Universal Pathogen Kit. 20 µL SYBR
qPCR were performed in triplicate on primers specific for the
target organism. Average of triplicate data shown.
12
FFPE DNA
FFPE DNA
MAG-BIND® FFPE DNA 96 KIT
qPCR FROM FFPE DNA
33
32
31
30
29
cT Value
The Mag-Bind® FFPE DNA 96 Kit is designed for the
rapid and reliable isolation of DNA from formalinfixed, paraffin-embedded (FFPE) tissue. The specially
formulated buffers reverse crosslinking and release short
and long DNA fragments. Mag-Bind® particles allow
for automation-friendly extraction from FFPE tissue in
under 6 hours. Purified DNA is suitable for downstream
applications including SNP analysis, sequencing and
genotyping.
This system is fully compatible on
Hamilton STAR®, Beckman Coulter Biomek® FX, Biomek®
2000, Tecan Fredom Evo®, Thermo KingFisher® Flex and
other workstations.
Omega Bio-tek Mag Bind
FFPE DNA
28
Company A Spin Column
27
26
25
24
Samples
Yield (µg/
section)
1
2
3
4
8.28
8.74
1.81
8.25
6.77
4.98
5.08
2.56
7.27
4.97
Averages
Company A
1
2
3
4
Averages
A260 / 280 A260 / 230
1.79
1.74
1.66
1.69
1.72
1.83
1.56
1.70
1.74
1.71
1.45
1.37
0.68
1.26
1.19
1.64
0.95
0.91
1.98
1.37
Comparison of Company A’s silica spin column
method for FFPE DNA purification versus Omega Biotek’s Mag-Bind® FFPE DNA Kit. Both protocols were
performed according to manufacturer’s recommened
protocols. Samples were quantitated using Thermo Fisher
NanoDrop® 2000c.
13
23
0.01
0.1
1
10
100
DNA (ng)
cT values of recovered DNA using the MagBind® FFPE DNA Kit and Company A Spin
Column Kit for FFPE DNA purification. A
10-fold dilution series of the recovered DNA
was used in a SYBR® Green-based real-time
PCR reaction targeting an 80-bp fragment of
the human ribosomal protein gene RPL13a.
Each reaction was performed in triplicate. The
input DNA amounts were 50, 5, 0.5, and 0.05 ng,
respectively.
Mag-Bind® FFPE DNA 96 Kit
Product
Preparations
M6958-00
1 x 96
M6958-01
4 x 96
FFPE RNA
MAG-BIND® FFPE RNA 96 KIT
The Mag-Bind® FFPE RNA 96 Kit is designed to isolate total RNA from formalin-fixed, paraffin-embedded
(FFPE) tissue samples. This kit uses Mag-Bind® technology for the efficient isolation of total RNA along with
optimized buffers that are designed to minimize the effects of formaldehyde modifications. Two options are
available for paraffin removal: a heat treatment or a xylene treatment. Samples can be processed manually or
on the Hamilton STAR and STARlet, Beckman Coulter Biomek® FX, Biomek® 2000, Tecan Freedom Evo®, Velocity
Biocell® and Thermo KingFisher® Flex workstations.
Source
125
MAG-BIND® FFPE RNA
96 KIT PROTOCOL
Lyse sample and
remove paraffin
Prepare lysate
Add Mag-Bind® Particles for
RNA binding and magnetize.
Aspirate supernatant
containing contaminants.
Sample
Type
Human
Breast
Tissue
Human
Kidney
Rat Liver
RNA Yield (µg)
Number
Company A
Omega
Bio-tek
1
2
3
4
1
2
1
2
3
4
5.8
12.1
9.4
5.4
5.7
9.1
28.3
15.1
26.7
8.2
13.9
30.1
26.4
18.3
28.4
26.5
42.8
28.6
32.5
9.4
Increase
%
139%
150%
181%
236%
402%
193%
51%
89%
22%
15%
RNA yield from various sample types using MagBind® FFPE RNA Kit versus Company A on Thermo
KingFisher® Flex. Both protocols were performed
according to manufacturer’s recommended
protocols. The Omega Bio-tek system provides better
sensitivity for gene expression profiling.
Wash Mag-Bind® Particles
Magnetize
Add DNase, wash,
and magnetize
Dry Mag-Bind® Particles
on magnet
Mag-Bind® FFPE RNA 96 Kit
Product
Preparations
M2551-00
1 x 96
M2551-01
4 x 96
Elute RNA
14
BLOOD DNA & RNA
E-Z 96® BLOOD & TISSUE DNA KIT
E-Z 96® Blood DNA Kit
Product
Preparations
D1196-00
1 x 96
D1196-01
4 x 96
12
10
DNA yield (μg)
The E-Z 96® Blood & Tissue DNA Kit can also be used for the
preparation of genomic DNA from buffy coat, serum,
plasma, saliva, cultured cells, buccal swabs and other
bodily fluids. An optimized lysis buffer and protease
digestion step removes nucleases, histones and other
proteins. After binding conditions are adjusted and DNA
is bound to the HiBind® matrix, the DNA is washed and
PCR inhibitors are removed. High quality DNA is eluted
and can be used for downstream applications such as PCR,
qPCR and microarray analysis.,
QUALITY & YIELD OF GENOMIC
DNA PURIFIED
8
6
4
2
0
Samples
2.0
1.9
A260/A280
Omega Bio-tek’s E-Z 96® Blood & Tissue DNA Kit provides
an easy and rapid method for the isolation of genomic,
mitochondrial or viral DNA. Up to 250 µL fresh, frozen or
anti-coagulated whole blood can be readily processed
at one time from common anticoagulants such as EDTA,
heparin and sodium citrate.
1.8
1.7
1.6
Samples
Quality and yield of genomic DNA purified using the E-Z 96® Blood
& Tissue DNA Kit. Purified genomic DNA from 200 µL whole human
blood was isolated with the E-Z 96® Blood & Tissue DNA Kit. DNA was
eluted in 100 µL and quantitated using Thermo Fisher NanoDrop 2000c.
MAG-BIND® PX BLOOD RNA 96 KIT
The Mag-Bind® PX Blood RNA 96 Kit provides a convenient and fast RNA extraction solution for samples
stabilized with PAXgene™ Blood RNA tubes or Applied Biosystems Tempus™ Blood RNA tubes. This system
combines Mag-Bind® particles and HiBind® silica technologies for purification of total RNA from up to 2.5 mL
preserved blood samples.
The blood sample is spun down and the crude RNA/DNA pellet is collected and washed. The pellet is then
resuspended and digested with proteinase K. The lysate is applied to a filter plate to remove genomic
DNA. This procedure completely removes inhibitors and sample stabilization reagents to allow for reliable
downstream analysis. High-quality purified RNA can be used for downstream applications such as qRT-PCR,
RT-PCR, and microarray analysis.
Mag-Bind® PX Blood RNA 96 Kit
Product
Preparations
M7763-00
1 x 96
M7763-01
4 x 96
15
TOTAL RNA
E-Z 96® TOTAL RNA KIT
RNA YIELD FROM TISSUE
50
40
RNA Yield (µg)
The E-Z 96® Total RNA Kit is designed for the
isolation of total cellular RNA from up to 5 x 106
cultured cells or soft tissues. This kit can process
single or multiple samples in less than 60 minutes.
Samples are lysed in a denaturing lysis buffer which
inactivates RNases. Binding conditions are adjusted
and then lysate is transferred to a 96-well HiBind® RNA
plate where the RNA is purified via three wash steps.
High-quality RNA is eluted in RNase-free water. RNA
purified using the E-Z 96® Total RNA method is ready
for applications such as RT-PCR, qPCR, differential
display, microarrays, and other downstream
applications.
30
20
10
0
E-Z 96® Total RNA Kit
Product
Preparations
R1034-00
1 x 96
R1034-01
4 x 96
R1034-02
12 x 96
Liver Kidney Spleen Heart Muscle Lung
Yield of isolated RNA with the E-Z 96® Total RNA
Kit. Purified total RNA from 20 mg various animal
tissues was isolated with the E-Z 96® Total RNA
Kit using a Thermo KingFisher® Flex. RNA was
quantitated using a Thermo Fisher NanoDrop®
2000c.
MAG-BIND® TOTAL RNA 96 KIT
RNA ISOLATION FROM SMALL
SAMPLE AMOUNTS
1500
1200
Yield (µg)
The Mag-Bind® Total RNA 96 Kit is designed for the
rapid and reliable isolation of total cellular RNA
from cultured cells and soft tissues. The system
utilizes the high nucleic acid binding properties of
Mag-Bind® particles resulting in increased sensitivity
over silica column-based systems. Mag-Bind® particles
are completely resuspended during the wash steps
to increase contact with wash buffers and enhance
removal of contaminants resulting in high-quality
RNA.
900
600
300
Mag-Bind® Total RNA 96 Kit
Product
Preparations
M6731-00
1 x 96
M6731-01
4 x 96
0
50K
10K
5K
1K
RNA Isolation from small sample amounts. Total RNA from
corresponding cell number amount HEK293 cultured cells
using the Mag-Bind® Total RNA 96 Kit. RNA was quantified
using Promega QuantiFluor®.
16
PLANT DNA
E-Z 96® PLANT DNA DS KIT
The E-Z 96® Plant DNA DS Kit is designed for the efficient recovery of genomic DNA up to 30 kb in size from
fresh, frozen or dried plant tissue samples rich in polysaccharides, polyphenols, or having a lower DNA
content. Up to 50 mg of wet tissue can be processed in less than 1 hour. The system combines the reversible
nuclec acid binding properties of the HiBind® matrix with the speed and versatility of spin column technology
to eliminate polysaccharides, phenolic compounds, and enzyme inhibitors from plant tissue lysates. Purified
DNA is suitable for PCR, restriction enzyme digestion and hybridization applications.
This procedure relies on the well-established properties of the cationic detergent, cetyltrimethyl
ammoniumbromide (CTAB), in conjunction with the unique binding system to increase yields and provide
high-quality DNA. The system eliminates the need for chloroform extractions traditionally associated with
CTAB-based lysis methods. Samples are homogenized and lysed in a high salt buffer containing CTAB and
binding conditions are adjusted and DNA is purified using a HiBind® DNA matrix. Salts, proteins, anad other
contaminants are removed to yield high-quality genomic DNA suitable for downstream applications such as
endonuclease digestion, thermal cycle amplification and hybridization applications.
COMPARISON OF DNA YIELDS
FROM MULTIPLE CROPS
150
Omega Bio-tek
Company Q
90
60
30
ac
co
To
b
n
ea
So
yb
n
Co
tto
n
Co
r
a
0
Ca
no
l
ng/mg
120
40-50 mg of respective fresh leaf tissue were
extracted in triplicate according to the manufacturer’s
recommended
protocol
and
eluted
in
100
µL.
DNA
analyzed
with
fluorescent
DNAbased quantfication method. Total yield was
divided by total tissue amount to show ng of DNA per
mg of leaf tissue.
17
E-Z 96® Plant DNA DS Kit
Product
Preparations
D1411-01
1 x 96
D1411-02
4 x 96
PLANT
MAG-BIND® PLANT DNA DS 96 KIT
The Mag-Bind® Plant DNA DS 96 Kit is designed for
difficult samples which are hard to lyse or contain
high amounts of polysaccharides and polyphenols.
This system uses a CTAB-based lysis buffer which
does not require organic solvents. The proprietary
binding system prevents inhibitors from binding to
the magnetic beads. The wash buffer system further
removes polysaccharides, phenolic compounds,
and enzyme inhibitors from plant tissue lysate. An
optional rebinding step is included with the kit,
but most samples do not require the additional
treatment.
Using paramagnetic particles provides highquality DNA that is suitable for direct use in most
downstream applications such as qPCR, PCR and
next generation sequencing.
The Mag-Bind® Plant DNA DS 96 Kit is compatible with
multiple liquid handlers and magnetic processors
such as Thermo KingFisher Flex, Qiagen BioSprint,
Applied Biosystems/Ambion MagMAX 96, Beckman
Coulter Biomek FX/NX, and Hamilton STAR/STARlet.
DNA YIELD COMPARISON
Type
Canola
Corn
Cotton
Potato
Soybean
Tobacco
Wheat
Omega
Bio-tek
59.0
29.8
63.5
206.5
25.4
19.4
152.3
Company Q
3.4
4.0
30.5
30.0
26.8
12.3
0.5
Approximately 50 mg leaf sample extracted per sample
according to manufacturer’s recommended protocols. DNA
concentration determined via flourescence-based nucleic acid
quantification. DNA quantification confirmed via SYBR qPCR (data not
shown). Amount of DNA per mg leaf sample shown above.
Mag-Bind® Plant DNA DS 96 Kit
Product
Preparations
M1130-00
1 x 96
M1130-01
4 x 96
E-Z 96® PLANT RNA KIT
The E-Z 96® Plant RNA Kit provides a convenient and rapid method for the isolation of total RNA from a
variety of plant samples. This kit provides an E-Z 96® Homogenizer Plate for the homogenization and
filtration of viscous plant cell lysates in combination with the HiBind RNA Plate for RNA purification. The
unique lysis buffer formulation contains chaotropic salts and detergents for efficient removal of contaminants
including polysaccharides and phenolic compounds. Purified RNA can be used for most downstream
applications such as RT-PCR, qPCR, Northern blot analysis, differential display, microarray analysis, whole
transcriptome sequencing and poly A+ RNA selection.
E-Z 96® Plant RNA Kit
Product
Preparations
R1027-01
2 x 96
R1027-02
8 x 96
Samples validated with the E-Z 96® Plant RNA Kit
Arabidopsis thaliana
Nicotiana tabacum
Beta vulgaris
Ornithogalum thyrsoides
Daucus carota
Oryza sativa
Euglena gracilis
Solanum tuberosum
Fragaria virginiana
Spinacia oleracea
Hordeum vulgare
Triticum aestivum
Humulus lupulus
Zea mays
Lycopersicon esculentum
18
400 Pinnacle Way Suite 450
Norcross, GA 30071
Phone: (800) 832-8896 / Fax: (770) 931-0230
info@omegabiotek.com
www.omegabiotek.com
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