1. science
2. biology
3. molecular biology

NUCLEIC ACID
ANALYSIS
Nucleic Acid Sample Preparation
Do you need pure RNA for QRT-PCR?
Is your DNA clean
and application-ready?
Do you want to skip
RNA isolation altogether?
SAMPLE PREPARATION REAGENTS >
Sample Preparation Reagents
Stratagene offers a full suite of nucleic acid isolation products to suit your
application. Our patented SideStep™ II QRT-PCR products allow you to skip
RNA purification altogether and proceed directly to a real-time QRT-PCR
amplification from cells, saving you valuable research time and money. This
method minimizes sample loss, making it ideal for high-throughput sample
processing and screening. Our highly popular, efficient and effective
Absolutely RNA® product line delivers intact, pure, high-quality total RNA and
mRNA for every application. These complete kits incorporate DNase treatment for the complete removal of genomic DNA. For DNA, we offer genomic
DNA purification kits that allow for a wide range of sample inputs, such as
tissue, cultured cells, and whole blood, and DNA extraction kits for PCR and
plasmid DNA isolation and purification.
CONTENTS:
1
2-3
4-5
6
7
Product Selection Guide
C e l l Ly s i s a n d R N A S t a b i l i z a t i o n
To t a l R N A P u r i f i c a t i o n
R N A f r o m P a r a f f i n - E m b e d d e d Ti s s u e
mRNA and miRNA Isolation
8
9
10
11
Genomic DNA Isolation
DNA Isolation from Gels, PCR
Plasmid DNA Isolation
Ordering Information
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Table 1
CHOOSE THE BEST SAMPLE PREPARATION PRODUCT FOR YOUR APPLICATION
NUCLEIC ACID TYPE
STRATAGENE SOLUTION
SAMPLE TYPE
SAMPLE SIZE
ADVANTAGES
RNA, DNA, miRNA
SideStep™ II QPCR and
QRT-PCR Products
Mammalian cells
Up to 1 x 10 6
+
+
+
+
Absolutely RNA®
Miniprep Kit
Mammalian cells
and tissue
10 5 to 10 7 cultured
cells or 5 to 40 mg
of tissue
+ Pure DNA-free total RNA for all downstream
applications
Absolutely RNA®
Microprep Kit
Mammalian cells
Up to 5 x 10 5 cells
+ Ideal kit for laser microdissection (LMD) samples
+ Single-spin cup protocol maximizes recovery of RNA
Absolutely RNA® 96
Microprep Kit
Mammalian cells
Up to 5 x 10 5 cells
+ High-throughput method purifies total RNA from
small RNA
Absolutely RNA®
Nanoprep Kit
Mammalian cells
1 cell to 1 x 10 4 cells
+ Small 10-μl elution volume for concentrated
RNA making it ideal for real time RT-PCR analysis
Absolutely RNA® FFPE Kit
Formalin-fixed paraffin
embedded tissues
(FFPE)
Variable
+ Reliable non-toxic method
+ Complete kit, no extra purchases required
+ Includes QPCR Reference RNA standard for gene
expression analysis
Absolutely mRNA™
Purification Kit
Total RNA
Up to 1 mg
+ Method delivers the purest mRNA
+ Fast 20-minute protocol
+ Scalable, simple kit method
SideStep™ mRNA
Enrichment Kit
Mammalian cells
Up to 1 x 10 6 cells
+ Delivers intact, highly pure mRNA directly from
cell culture
+ Swift 30-minute protocol
+ Scalable, simple kit method
SideStep ™ Lysis and
Stabilization Buffer
Mammalian cells
Up to 1 x 10 6
+
+
+
+
DNA Extraction Kit
Mammalian tissue,
cell, or blood
Up to 1 x 10 8 cells;
250 mg tissue
+ Easy-to-use, scalable, non-toxic
method for DNA extraction
DNA Micro Extraction Kit
Blood
100 – 300 ml blood
+ Highly reliable, rapid 60-minute protocol
High Molecular
Weight Genomic DNA
Recoverease ™ DNA
Isolation Kit
Whole mammalian
tissues: liver, spleen,
kidney, brain, and
testes
40 mg to 185 mg
mammalian tissue
+ Recover high-molecular-weight (100 to 500 kb)
genomic DNA fully hydrated and ready for multiple
downstream applications
+ Non-toxic method
+ No organic extractions or ethanol precipitations
PCR Product
Clean-up
StrataPrep ® Gel
Extraction Kit
Agarose gel slices
Variable
+ Effective purification of DNA fragments from 100 bp to
greater than 23 kb
+ Versatile protocol—works with both conventional and
low-melt agarose gels
StrataPrep ® PCR and 96
PCR Purification Kits
PCR reaction
mixtures
50- 100 μl
+ Rapid, high-yield PCR product purification up to 10 μg
StrataPrep ® Plasmid
Miniprep Kit
Bacterial cultures
1.0 – 3.0 ml cultures
+ Rapid, phenol-free method for purifying pure
plasmid DNA
+ Accommodates all sizes of DNA from 2.9-kb
plasmids to 45-kb cosmids
StrataPrep ® EF Plasmid
Midi Prep Kit
Bacterial cultures
25 to 200 ml cultures
depending on
plasmid copy number
+ Rapid, high yields of pure endotoxin-free DNA
+ Improves transfection efficiency
+ Purification of DNA ranging from 2.9-kb plasmids
Total RNA
Messenger RNA
Genomic DNA
Plasmid DNA
Complete RNA transcriptome
Single-tube format eliminates sample loss
Go directly to QRT-PCR analysis
Fast 10-minute protocol
Single-tube format eliminates sample loss
Go directly to QPCR from cells
Non-toxic phenol-free method
Fast 10-minute protocol
to 48-kb cosmids
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2>3
CELL LYSIS AND RNA STABILIZATION > SIDESTEP ™ II QRT-PCR PRODUCTS
Obtain RNA and DNA Directly from
Cells without Loss
The SideStep™ II QPCR and QRT-PCR products allow you to quantitate gene expression directly
from cells. Our kits deliver efficient cell lysis, RNA stabilization, gDNA removal and quantitative
gene expression analysis—without the need for a separate RNA purification step. Our SideStep™
kits employ a patent-pending solution that lyses cells and immediately stabilizes the released
nucleic acids for many months. The SideStep™ method allows you to completely avoid nucleic
acid purification and proceed directly to cDNA synthesis and real-time QPCR detection.
SideStep ™ Lysis and Stabilization Process
Our SideStep™ Lysis and Stabilization Buffera lyses cells in 10
minutes at room temperature, with the actual lysis step taking
only 1 minute (Figure 1). RNA, miRNA, and DNA are released
from the cells and immediately stabilized. RNA remains intact
and high quality even after 20 months of -80°C storage in the
lysate (Figure 2). The SideStep buffer offers the best cell lysis
and is easier to use than other cell lysis methods. Our
standard SideStep protocol permits lysis of up to 1 million
Conventional Method
SideStep™ Method
Cells
Cells
Lysis
Lysis
cells in 100 μ l and is scalable.
QRT-PCR Normalization Using Genomic DNA
Our Quantos QPCR normalization primers, included in all
SideStep II kits, are unique single-copy sequences on different
human chromosomes which provide a standard for evaluation
of gDNA amount. Additionally, they are useful for determining
cell number in an unknown sample and determining relative or
comparative quantification of mRNA copy number in multiple
(Optional
DNase Treatment)
samples using genomic DNA as the normalizer.
Bind to Column Matrix
QRT-PCR
DNase Treatment
2 Steps > 10 Minutes
Elute
QRT-PCR
6 Steps > 30 Minutes
Figure 1
S IDE S TEP ™ PRODUCTS SAVE TIME, INCREASE THROUGHPUT
Our SideStep™ product line takes you directly from cells to QRT-PCR in fewer steps
with less time, increasing your throughput.
cDNA Synthesis Directly from Cells
Our SideStep™ II QPCR cDNA Synthesis Kita employs direct synthesis of cDNA
from cells without having to first isolate RNA. This highly sensitive and reliable
method takes only 15 minutes to perform using SideStep cell lysates and
allows you to generate cDNA expressly for QPCR analysis. This high sensitivity
is due to our QPCR-grade and nuclease-free AffinityScript™ Multiple
Temperature Reverse Transcriptasea as well as a highly efficient cDNA synthesis buffer. Additionally, the cDNA synthesis reagents are formatted in a convenient master mix for speed and high reliability.
Multiple Formats Offer SideStep™ Technology
Rapid and precise real-time analysis directly from cell lysates is now possible
with our SideStep™ II QPCR and QRT-PCR Kitsa,b. Our application-tested kits
employ detection of RNA using both one-step and two-step formats (Table 2).
Ideally, RT-PCR is performed using primers designed to span exon-exon junctions, however, we include genomic DNA removal reagents in all Sidestep II
kits. Genomic DNA, miRNA, and RNA can all be measured directly from the
SideStep lysates. RNA is stable in the lysate for up to 20 months at -80°C,
which is ideal for a host of cell-based gene expression applications such as
mRNA profiling and siRNA knockdown validation.
Figure 2
S IDE S TEP ™ CELL LYSATES STABLE AT LEAST 20 MONTHS AT -80°C
We purified total RNA from a lysate stored at -80°C for 20 months and analyzed it
using an Agilent® bioanalyzer. Our data clearly show no degradation of RNA indicated
by sharp peaks for the 28S and 18S ribosomal bands and a RIN score of 8.7.
SIDESTEP™ PRODUCTS
APPLICATION
KEY ADVANTAGES
SideStep™ II Cell Lysis Analysis Kit
Cell lysate analysis
+ Analysis of RNA, DNA, miRNA, mRNA and proteins
+ Includes genomic DNA removal reagents
+ Includes Quantos QPCR normalization primers for
comparative quantification or cell number determination
SideStep™ II QPCR cDNA Synthesis Kit
cDNA synthesis
from cell lysates
+ Go from cells to cDNA in 25 minutes
+ Includes AffinityScript™ Multiple Temperature RT for high
yields of full length cDNA
+ Includes genomic DNA removal reagents prior to cDNA synthesis
SideStep™ SYBR® Green QPCR Master Mix
QPCR from cell
lysates
+ DNA quantification directly from cell lysates
SideStep™ II QRT-PCR Master Mix, 1-Step (probe)
QRT-PCR from
cell lysates
+ Analysis of RNA, DNA, miRNA, and mRNA
SideStep™ II QRT-PCR Master Mix, 2-Step (probe)
QRT-PCR from
cell lysates
+ Analysis of RNA, DNA, miRNA, and mRNA
SideStep™ II SYBR® Green QRT-PCR Master Mix, 2-Step
QRT-PCR from
cell lysates
+ Analysis of RNA, DNA, miRNA, and mRNA
SideStep™ mRNA Enrichment Kit
mRNA isolation
directly from cell
lysates
+ Pure mRNA directly from cells in 30 minutes
Table 2
CHOOSE THE RIGHT S IDE S TEP ™ PRODUCT FOR YOUR RESEARCH
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4>5
TOTAL RNA PURIFICATION > ABSOLUTELY RNA ® KITS
High Yields of Pure DNA-Free Total RNA
Our Absolutely RNA® Kits offer easy-to-use methods that deliver high yields of intact, full-length
DNA-free RNA that can be used for a variety of downstream applications. Additionally, our kits
accommodate a wide range of sample types and amounts, allowing maximum flexibility.
Highly Pure and DNA-Free
High-Quality RNA, Convenience, and Savings
RNA quality and purity are paramount for successful Northern
Our Absolutely RNA isolation kits contain all of the necessary
blot analysis, reverse-transcriptase PCR (RT-PCR), microarray
reagents you need (including DNase I) for isolating pure total
target labeling, or QRT-PCR experiments. Carryover of phenol,
RNA, thus saving you money and time. Additionally, our DNase
DNA contamination, and RNA degradation can contribute to
is supplied lyophilized, making our kits perfect for room temper-
poor labeling and signal quality in microarray experiments and
ature storage and saving you valuable freezer space. Removal of
inhibit RT-PCR or QRT-PCR amplification. Using clean, DNA-
genomic DNA is recommended for sensitive gene expression
free, total RNA improves detection and discrimination of low
studies where microarray or QRT-PCR techniques are employed.
expression level genes in virtually every application.
Using our Absolutely RNA kits gives you flexibility in primer
design, since you don’t have to worry about genomic DNA
Fast, Easy, Non-Toxic Method
Our Absolutely RNA kits use a safe, rapid, phenol-free method
®
that works with tissue or cultured cells. The method for all kits
carrythrough. We also offer a variety of Absolutely RNA kits to
suit your application or sample size (Table 3).
is simple. Isolation of total RNA is performed in a single spin
cup to minimize potential loss of RNA during the procedure.
Following lysis, the solution is applied to the silica-based fiber
matrix within the spin cup to bind the RNA. Genomic DNA
contamination is effectively removed by incubation with DNase I.
The RNA is then washed to remove contaminants and total RNA
is recovered in the appropriate final elution buffer (Figure 3).
Figure 3
ABSOLUTELY RNA ® KIT METHOD
Sample Size
Format
Capacity
Elution Volume
Application
ABSOLUTELY RNA ®
MINIPREP KIT
ABSOLUTELY RNA ®
MICROPREP KIT
ABSOLUTELY RNA ®
96 MICROPREP KIT
ABSOLUTELY RNA ®
NANOPREP KIT
ABSOLUTELY RNA ®
FFPE KIT
ABSOLUTELY mRNA ™ KIT
105 to 107 cells
or 5-40 mg of tissue
Up to 5 x 105 cells
Up to 5 x 105 cells
1 x 104 cells down
to a single cell
Formalin-fixed paraffinembedded (FFPE) tissues
Up to 1 mg of total RNA
Spin cup
Spin cup
Spin cup
Spin cup
Spin cup
Precision engineered
oligo-dT magnetic particles
> 200 μg RNA
50 μg RNA
50 μg RNA
Approx. 200 ng RNA
50 μg RNA
1 mg of total RNA/ 1 mg of
magnetic particles
50 μl
30 μl
30 μl
10 μl
30 μl
10-50 μl
RT-PCR, QRT-PCR,
Northerns, microarray
target generation
LMD, QRT-PCR,
microarray target
generation
High-throughput
QRT-PCR and
microarray target
generation
QRT-PCR, LMD
QRT-PCR, LMD
RT-PCR, QRT-PCR,
Northerns, microarray target
Table 3
CHOOSE THE BEST KIT FOR YOUR APPLICATION
TOTAL RNA PURIFICATION > MULTIPLE ABSOLUTELY RNA ® KIT FORMATS
Multiple Kit Formats Accommodate
Wide Sample Size Range
Flexible Starting Amounts of Cells or Tissue
High Throughput Total RNA Purification
Our Absolutely RNA Miniprep Kit is the only kit that produces high yields of
Our Absolutely RNA 96 Microprep Kit efficiently isolates high-quality
total RNA for RT-PCR, QRT-PCR, microarray target samples, Northern blot
total RNA from a variety of cultured cells from 10 cells up to 5 x 10 cells.
®
®
5
analysis, and in vitro transcription reactions. Our kit methodology accommo-
This kit is perfect for RNA purification for all of your high-throughput
dates a wide range of tissue and cell amounts without compromising RNA
purification needs. The purified RNA can be used for RT-PCR, QRT-PCR,
yield or integrity and coupled with an effortless on-column DNA removal
and expression analysis studies.
step gives you pure, DNA-free RNA from cells and tissues (Figure 4).
Single Cell RNA Isolation
Optimized For Cultured Cells and Laser Microdissection (LMD)
Our Absolutely RNA Nanoprep Kit is the only kit qualified to isolate
Our Absolutely RNA Microprep Kit isolates total RNA from small starting
RNA from a single cell (Figure 5). Total RNA yields are high-quality and
samples and LMD samples. This kit is designed for samples containing up
concentrated in a small 10-μl elution volume which makes it the perfect
®
5
to 5 x 10 cells. Isolation of total RNA takes place in a single spin cup, with
®
kit for QRT-PCR.
DNA removal performed directly in the spin cup to minimize potential loss
of RNA during the procedure. The resulting RNA is ideal for QRT-PCR,
microarray target generation, and other applications.
104
103
5 x 103
Fluorescence (dR)
4 x 103
3 x 103
102
2 x103
1 x 10
10
1
3
0 x 10
2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40
Cycles
Figure 4
Figure 5
EXCEPTIONAL TOTAL RNA PURITY
ABSOLUTELY RELIABLE RNA PURIFICATION DOWN TO A SINGLE CELL
We assessed the purity of a total RNA sample isolated from a brain cell line using the
Absolutely RNA® Kit method with an Agilent® bioanalyzer. An RNA ratio (28S/18S) greater than
1.5 indicates very pure RNA. The data shown here yielded an RNA ratio 1.72, demonstrating
the recovery of very pure RNA using the Absolutely RNA® Kit method.
QRT-PCR results using total RNA template isolated from 104, 103, 102, 10 and 1 HeLa cell.
Duplicate reactions were run for each sample with human GAPDH molecular beacon and
primers and 2 μl RNA template.
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RNA FROM PARAFFIN-EMBEDDED TISSUE > ABSOLUTELY RNA ® FFPE KITS
QPCR-Grade RNA from ParaffinEmbedded Tissues
The Absolutely RNA® FFPE Kit delivers a reliable, easy-to-use, non-toxic method to isolate
quality RNA from formalin-fixed paraffin embedded (FFPE) tissue sections and includes a quantitative RNA reference standard for meaningful QRT-PCR expression analysis.
Reliable RNA Recovery from FFPE Tissue
Easily Evaluate and Cross Compare Data
Our Absolutely RNA FFPE Kit allows you to purify QRT-PCR-
There can be a high degree of variability between blocks of
grade total RNA from formalin-fixed paraffin-embedded tissues.
tissue, even in 10-micron sections of paraffin-embedded tissue.
The kit contains optimized reagents for improving the reliability
This may be due to differences in tissue harvest protocols, the
of RNA recovery from these difficult-to-use samples. Using
nature of the tissue, fixation procedures or the age or storage
traditional methods, Proteinase K digestion is either inefficient
condition of this archived material. To minimize this variability,
or ineffective at reversing the RNA modifications caused by the
we include our Stratagene QPCR Human Reference RNA to use
fixation process, resulting in poor and unreliable RNA recovery.
as a baseline to compare gene expression from experiment to
In contrast, our method using an optimized in-solution
experiment. In addition, every lot of our kit is application tested
Proteinase K digestion, which completely reverses the RNA
and qualified using QRT-PCR, ensuring that you will recover
modifications, maximizes RNA recovery and minimizes your
the maximum amount of total RNA from FFPE tissues.
®
®
time spent during this procedure. As a result, our kit produces
more RNA suitable for QRT-PCR (Figure 6) and delivers higher
sensitivity in QRT-PCR (Table 4) compared to a leading competitor.
10,000
8,000
Total RNA (ng)
6>7
6,000
C t VALUE
STRATAGENE
COMPANY A
STRATAGENE
COMPANY A
4,000
Target
ß2M
ß2M
GAPDH
GAPDH
2,000
Colon 3 hr digestion
20.58
24.99
27.83
33.41
Liver 3 hr digestion
24.79
27.67
31.16
36.57
0,000
Liver 3 hr.
Absolutely RNA® FFPE Kit
Liver 18 hr.
Company A RNA FFPE Kit
Figure 6
Table 4
MORE YIELD IN LESS TIME
MORE QRT-PCR GRADE RNA
Our Absolutely RNA® FFPE Kit resulted in better RNA yield compared to
Company A's kit, even after using an 18-hour Proteinase K digestion (which
can help with difficult tissues).
The Absolutely RNA® FFPE Kit delivers more RNA available for QRT-PCR analysis resulting in
better sensitivity than RNA isolated using an FFPE kit from Company A. We performed 1-step probebased QRT-PCR amplifications on two targets using 100 ng of RNA isolated from the Absolutely
RNA® FFPE Kit and Company A. Using RNA from the Absolutely RNA® FFPE Kit produced much
lower Ct values on both RNA targets and tissue types. (ß2M = Beta-2-microglobulin)
mRNA AND miRNA ISOLATION > ABSOLUTELY mRNA ™ , SIDESTEP ™ , miRACLE ™ KITS
Highly Pure mRNA and miRNA in Minutes
Our Absolutely mRNA Purification Kit maximizes mRNA yield and purity from total RNA. Our
SideStep mRNA Enrichment Kit provides the perfect method to isolate clean mRNA directly from
cultured cells. Removal of predominant larger RNAs is required for accurate analysis of miRNA
expression by QPCR. The miRACLE miRNA Isolation kit purifies small miRNAs with minimal
contamination from large RNA molecules or genomic DNA.
™
™
™
Maximize Yield, Minimize Contamination
Pure mRNA Directly from Cells
Our Absolutely mRNA Purification Kit produces high yields of
Our SideStep mRNA enrichment kita allows you to purify mRNA
pure mRNA directly from RNA using a 20-minute protocol.
directly from cells using an exceptionally efficient 30-minute
The kit includes customized oligo-dT magnetic particles that
protocol. Using the SideStep technology, cell lysis and nucleic acid
exclusively bind mRNA to maximize yield and quality and mini-
stabilization are performed in the same buffer, ensuring maximum
mize rRNA contamination (Figure 7). We selected an oligo-dT
recovery of mRNA while eliminating the steps for total RNA purifica-
magnetic bead technology because it offers exceptional binding
tion. Pairing this with our Absolutely mRNA technology, you recover
capacity and specificity. Furthermore, older generation magnetic
the highest yields of pure mRNA, useable for every downstream
bead isolation kits sometimes resulted in leaching of magnetite
application, including microarray target labeling and QRT-PCR
and thus the recovery of ‘dark colored’ RNA product. The
(Figure 8).
™
oligo-dT magentic beads in our kit have a reticulated outer shell
constructed of a synthetic polymer coating, which confines the
MicroRNA from Cells and Tissue
magnetite within each bead and prevents leaching.
Our miRACLE miRNA Isolation Kit offers a fast and highly efficient
™
method for miRNA isolation. The kit employs acidified phenol and
chloroform to select for RNA molecules and a specially designed
column for enrichment of small miRNAs. Our miRACLE miRNA isolation kit removes genomic DNA and large RNA molecules from each
preparation, ensuring excellent sensitivity in downstream applications
such as QRT-PCR and miRNA microarray analysis.
0.6
mRNA
Immobilization
Contaminant
Removal
Flourescence (dRn)
0.5
mRNA
Elution
0.4
100 cells
0.3
10 cells
1 cell
0.2
mRNA
contaminant
0.1
magnetic bead
0.0
10
20
30
40
50
Cycles
Figure 8
Figure 7
ABSOLUTELY mRNA ™ KIT PURIFICATION METHOD
™
Our Absolutely mRNA Purification Kit uses a 3-step protocol to produce pure,
contaminant-free messenger RNA in 20 minutes.
OBTAIN QRT-PCR GRADE mRNA USING S IDE S TEP ™ mRNA ENRICHMENT KIT
We isolated mRNA from 100, 10, 1 cell equivalents (blue lines) using the SideStep™ mRNA
enrichment kit. Our data clearly shows sensitive and linear QRT-PCR amplification of GAPDH
down to a single cell. The reaction efficiency was 91% and Rsq = 1.00. Reactions were run on the
Mx3000P® QPCR System.
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8>9>10
GENOMIC DNA ISOLATION > DNA EXTRACTION KITS AND RECOVEREASE ™ DNA ISOLATION KITS
Simple, Non-Toxic Methods Deliver
Pure Genomic DNA
Our DNA Extraction Kits and Recoverease DNA Isolation Kit deliver pure intact genomic DNA
from blood, cells, or tissue.
™
Scalable DNA Extraction Method
Easily Recover High Molecular Weight DNA
Our DNA Extraction and DNA MicroExtraction Kits use a non-
Our RecoverEase DNA Isolation Kit recovers high molecular
toxic method to rapidly and reliably isolate high molecular weight,
weight (100-500 kb) genomic DNA from whole tissues such as
PCR-ready genomic DNA from whole blood samples as large as
liver, spleen, kidney, brain, and testes without hazardous and
5 ml, or as small as 1 μl, respectively. Using a modification of the
tedious organic extraction or ethanol precipitations. The entire
™
1
protocol developed by Miller, et. al. , separation of the proteins is
purification procedure requires less than one hour of hands-on
accomplished by a “salting out” procedure, followed by ethanol
time and one overnight incubation so the DNA is ready for use
precipitation of DNA and resuspension in your buffer of choice.
the next day (Figure 9).
The number of samples that can be processed simultaneously
with this technique is limited only by the amount of centrifuge
space available. Additionally, our DNA Extraction kit can be used
to isolate DNA from a variety of sample sources (Table 5).
RecoverEase™
Method
Phenol-Chloroform and
Ethanol Method
Tissue Disaggregation
(3 minutes/tissue)
SOURCE
QUANTITY
YIELD
SIZE (kb)
TIME
Whole Blood
5 ml
> 100-300 μg
100 - 500
2 hours
Proteinase Digestion
(3 hours)
Phenol-Chloroform
Extraction
(35 minutes/tissue)
Solid Tissue
1 gm
> 250 μg
50 - 100
2 hours, 45 minutes
Tissue-Culture Cells
108 cells
> 600 μg
50 - 100
2 hours, 15 minutes
Ethanol Precipitation
(10 minutes/tissue)
DNA Rehydration
( 48 - 72 hours)
Total Time: > 3 days
Coarse Filtration
(30 seconds)
Nuclei Isolation
(15 minutes)
Proteinase Digestion
(45 minutes)
Dialysis and Digestion
(16 hours)
Total Time: 17.1 hours
Table 5
FIGURE 9
OUR FLEXIBLE KIT CAN BE USED TO ISOLATE DNA FROM TISSUE, CELLS, OR
WHOLE BLOOD
SAVE TIME USING R ECOVER E ASE ™ DNA ISOLATION KIT
FOR GENOMIC DNA PURIFICATION
Our DNA Extraction Kit uses a non-toxic method to efficiently purify high yields of genomic DNA.
DNA ISOLATION FROM GELS, PCR, PLASMIDS > STRATAPREP ® DNA PURIFICATION KITS
Novel Purification Methods for a
Variety of Applications
StrataPrep® DNA Purification Kits utilize a special silica-based fiber matrix technology in a microspincolumn format to capture DNA that is application-ready in minutes. There is no need for aliquotting
messy slurries, extracting with phenol, or concentrating the DNA via alcohol precipitations.
Pure DNA from Agarose Gels
Endoxin-Free Plasmid DNA for Transfection
Our StrataPrep DNA Gel Extraction Kit allows rapid, high-yield isola-
Our StrataPrep EF Plasmid Midiprep Kit is a fast and easy way to isolate
tion of pure DNA from agarose gels. The DNA extraction solution is
up to 350 μg of endotoxin-free plasmid DNA. The presence of endotox-
added directly to the gel slice of interest and then transferred to the
ins (also known as lipopolysaccharides) sharply reduces transfection
unique microspin cup. Once bound, the DNA is washed and eluted
efficiencies in sensitive cell culture lines. The StrataPrep EF plasmid
from the column and is ready for use.
midiprep kit includes a unique and easy optional color indicator method
®
®
to effectively reduce endotoxin to levels of less than 0.1 EU (endotoxin
Fast, Effective Purification Method for PCR Products
units) per μg DNA, which significantly improves your transfection effi-
Our StrataPrep PCR and StrataPrep 96 PCR Purification Kits prepare
®
®
PCR products quickly and easily for downstream applications. The DNA
binding solution is added directly to the PCR microcentrifuge tube or 96-
ciencies (Figure 12). The convenient protocol reliably yields DNA that is
pure and ready for the most rigorous downstream applications.
well plate and transferred to a microspin cup or 96-well binding plate
Rapid, Small Scale Plasmid DNA Purification
containing a specialized silica-based fiber matrix. Once bound, the DNA
Our StrataPrep Plasmid Miniprep Kit provides a rapid, phenol-free
is washed, eluted, and ready for use. The StrataPrep 96 PCR purification
method for purifying pure plasmid DNA. The bacteria is pelleted, lysed,
kit effectively purifies a range of PCR products for downstream applica-
and then neutralized. The pre-cleared lysate is mixed with the DNA
tions such as microarray spotting, sequencing, and cloning (Figures 10
binding solution and transferred to the micro-spin cup. Once bound, the
and 11).
DNA is washed, eluted, and ready for use in virtually any downstream
®
application.
Yield
DNA Size
Minimum Elution Volume
StrataPrep® Gel
Extraction Kit
StrataPrep® PCR
Purification Kit
StrataPrep® 96 PCR
Purification Kit
StrataPrep® Plasmid
Miniprep Kit
StrataPrep® EF
Midiprep Kit
10 μg
10 μg
10 μg
20 μg
350 μg
100 bp to 23 kb
100 bp to long PCR
amplicons
100 bp to long PCR
amplicons
2.9-kb plasmids to
45-kb cosmids
2.9-kb plasmids to
48-kb cosmids
50 μl
50 μl
50 μl
50 μl
0.5 ml
Table 6
CHOOSE THE BEST S TRATA P REP ® KIT FOR YOUR DNA ISOLATION AND PURIFICATION NEEDS
DNA ISOLATION FROM GELS, PCR, PLASMIDS > STRATAPREP ® DNA PURIFICATION KITS
Figure 10
OBTAIN HIGH QUALITY SEQUENCE DATA FROM DNA PURIFIED WITH OUR S TRATA P REP ® KITS
% ß-galactosidase expression
100
kb
80
60
40
20
0
StrataPrep® EF Plasmid Midiprep Kit
Competitor's endotoxin-free system
Figure 11
Figure 12
OUR S TRATA P REP ® KIT EFFECTIVELY PURIFIES A WIDE SIZE
RANGE OF DNA
ACHIEVE HIGH TRANSFECTION EFFICIENCY USING OUR
ENDOTOXIN-FREE S TRATA P REP ® PLASMID DNA
Our StrataPrep® PCR Purification Kit delivers highly pure PCR-generated DNA
fragments ranging from 250 bp to 23 kb using a fast and easy method.
pCMV-ß-gal plasmid DNA was prepared with our StrataPrep® EF Plasmid
Midiprep Kit and a competitor’s endonuclease-free plasmid purification system.
Cos-1 and PC-3 cell lines were transfected by both a calcium phosphate (CP)
method and our LipoTaxi® transfection reagent.
www.stratagene.com
Ordering Information
DESCRIPTION
SIZE
CATALOG NO.
For Direct to QRT-PCR from Cell Lysates
SideStep™ SYBR® Green QPCR Master Mix
400 reactions
400904
S i d e S t e p ™ I I C e l l Ly s a t e A n a l y s i s K i t
100 lysis reactions
400916
SideStep™ II QPCR cDNA Synthesis Kit
50 reactions
400908
S i d e S t e p ™ I I Q RT- P C R M a s t e r M i x K i t , 1 - s t e p
400 reactions
400917
S i d e S t e p ™ I I Q RT- P C R M a s t e r M i x K i t , 2 - s t e p
400 reactions
400918
S i d e S t e p ™ I I S Y B R ® G r e e n Q RT- P C R M a s t e r M i x K i t , 2 - s t e p
400 reactions
400909
Total RNA Purification from Paraffin-Embedded Tissues
Absolutely RNA® FFPE Kit
50 preps
400809
Absolutely RNA® FFPE Kit (w/o deparaffinization reagents)
50 preps
400811
Absolutely RNA® Miniprep Kit
50 preps
400800
Absolutely RNA® Microprep Kit
50 preps
400805
Absolutely RNA® 96 Microprep Kit
2 plates
400793
10 plates
400794
50 preps
400753
Total RNA Purification
Absolutely RNA® Nanoprep Kit
Messenger RNA Isolation
Absolutely mRNA™ Purification Kit
10 preps
400806
SideStep™ mRNA Enrichment Kit
10 preps
400902
miRACLE™ miRNA Isolation Kit
50 preps
400813
miRACLE™ miRNA Isolation Kit w/o Phenol/Chloroform
50 preps
400815
MicroRNA Isolation
PCR DNA Isolation
StrataPrep® Gel Extraction Kit
StrataPrep® PCR Purification Kit
StrataPrep® 96 PCR Purification Kit
50 preps
400766
250 preps
400768
50 preps
400771
250 preps
400773
2-plate kit
400775
10-plate kit
400776
Plasmid DNA Isolation
StrataPrep® EF Midiprep Kit
StrataPrep® Plasmid Miniprep Kit
20 preps
400721
100 preps
400722
50 preps
400761
250 preps
400763
Genomic DNA Isolation
DNA Extraction Kit
35 extractions
200600
DNA MicroExtraction Kit
100 extractions
200601
RecoverEase™ DNA Isolation Kit
30 preparations
720202
15 preparations
720203
Related Products
Endonuclease A Removal Buffer
500 ml
400723
R N A Ly s i s B u f f e r
125 ml
400792
LEGAL LANGUAGE
®
REFERENCES
®
®
Absolutely RNA , Mx3000P , and StrataPrep are registered trademarks of Stratagene in the United States.
Absolutely mRNA , miRACLE , RecoverEase , and SideStep are trademarks of Stratagene in the United States.
™
™
™
™
®
LipoTaxi is a registered trademark of Genetic Applications LLC.
SYBR is a registered trademark of Molecular Probes.
®
a. Patents pending.
b. Patents pending. Purchase of this product is accompanied by a license under the foreign counterparts of U.S. Patents
Nos. 4,683,202, 4,683,195 and 4,965,188 for use in the polymerase chain reaction (PCR) process, where such process
is covered by patents, in conjunction with a thermal cycler whose use in the automated performance of the PCR process
is covered by the up-front license fee, either by payment to Applied Biosystems or as purchased, i.e., an authorized
thermal cycler.
1. Miller, S.A., Dykes, D.D., and Polesky, H.F. ( 1988), Nucleic Acids Res. 16: 1215
A M P L I F I C AT I O N
CELL BIOLOGY
CLONING
M I C R O A R R AY S
NUCLEIC ACID
A N A LY S I S
PROTEIN FUNCTION
& ANALYSIS
Q U A N T I TAT I V E
PCR
S O F T WA R E
SOLUTIONS
Ordering Information:
Find an Agilent customer center in your country:
www.agilent.com/chem/contactus
U.S. and Canada:
1-800-227-9770
[email protected]
Asia Pacific:
[email protected]
Europe:
[email protected]
This information is subject to change without notice.
© Agilent Technologies, Inc. 2008
Printed in USA, August 2008
5989-8236EN
BR67-04/07 (71035)