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GE Healthcare Life Sciences

Detection Sample collection

Clean-up

Amplification Isolation and purifcation

Sample handling and preparation within the genomics workflow

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Sample handling and preparation within the genomics workflow Contents Introduction

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Chapter 1

Dried biosample collection with Whatman™ FTA™ cards

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Chapter 2

Ready-To-Go ™ and amplification techniques

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Chapter 3

Fluorescent labelling and detection of biomolecules

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Introduction Welcome to our eBook, we hope you find it enjoyable and informative.

Across three chapters, the eBook introduces some key sample handling and preparation stages within the genomics workflow. Particular attention is given to sample collection, ambient temperature stabilization, amplification, and fluorescent labelling and detection.

Each chapter contains useful practical information and guidelines providing a unique insight for those involved in genomics research and assay development.

To take part in the discussion visit our blog at: www.gelifesciences.com/dxcomponents And follow us on Twitter at: @GEdiagnostics

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Chapter 1

Dried biosample collection with Whatman FTA cards

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The following are important considerations during biosample collection a) b) c)

Collection Transportation Storage

d)

Processing

Current practices include: Specialist collection Transport in dry ice/wet ice Refrigerated, freezer or cryogenic storage Downstream processing for DNA, protein and metabolite analysis

a)

b)

c)

d)

This cold chain logistical workflow, which can be expensive both to establish and maintain, is essential for biosamples where maintaining a sample’s integrity over time is a key requirement. 5


Many issues associated with the transportation, stabilization, and storage of biological samples can be alleviated by using dried biosample collection.

Dried biosample collection Biological samples such as blood, serum, buccal cells, urine, or tissue homogenate are transferred to a paper substrate and subsequently dried. The biological molecules of interest within the sample are then extracted for subsequent analysis.

Dried blood spot (DBS) collection has emerged as a useful, alternative method to traditionally established wet collection. It has been widely employed in screening programs following the publication of its use in the detection of phenylketonuria (PKU) in large populations of newborn infants(1).

6 1.

Guthrie and Susie, Pediatrics 32:3 338-343 (1963).


Safer Reduces risk by improving the security of supply and handling

Future The technique is suited to changing technology and user/patient needs

Dried biosample collection

Easier Simplified workflow supports collection, transport and storage in remote and challenging environments

Leaner Reduces cost and removes steps from an existing wet sample collection workflow

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We offer a broad portfolio for the collection and ambient temperature transport and storage of a range of biosamples.

Whatman FTA cards

Chemically coated matrices provide long-term stability of DNA at ambient temperature*.

Lyse cells on sample application Simplified biosample collection workflow No need for refrigeration and cold chain logistics

* DNA analysis performed on 22 year-old blood and 12 year-old buccal samples.

903™ collection cards

Proven for blood collection and widely used in neonatal screening programs.

CE marked for the collection of blood samples from neonates and adults by healthcare professionals.

FDA-registered in vitro Class II Medical Device Compliant with Directive 98/79/EC CE marked as an in vitro Diagnostic Medical Device in Europe 903 Protein Saver Card is a "research use only" format.

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Whatman FTA cards: two different chemistries

Whatman FTA chemistries

Whatman FTA: use when DNA needs to be associated with the card/punch DNA remains tightly bound to the matrix while cell membranes and organelles are lysed and proteins and inhibitors are washed away • • •

Whatman FTA Elute: use when DNA is required in solution Yields pure DNA in solution, with a simple protocol which eliminates the need for a DNA purification kit Whatman FTA

Whatman FTA Elute

Collect sample and capture DNA in one step Purify DNA in less than 30 min Compatible with a wide range of samples

• Compatible with downstream applications such as Q-PCR. • Supports small sample volume needs (12 to 125 µl) • DNA in 15 to 30 min

DNA remains on card Associate & Store

Yields DNA in solution Rapid analysis

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Which is the correct card for the collection of DNA?

What is your biological sample? Coloured

Plant

Clone

FTA Plant Saver

FTA CloneSaver™

Is DNA required in solution or associated with card?

Is DNA required in solution or associated with card?

With card

In solution

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FTA Micro

2

FTA Mini

With card

In solution

Number of sample spots per card

FTA Elute Micro

Colourless

Number of sample spots per card

IND FTA Elute Micro

1

4

FTA Classic

FTA Micro

2

4

FTA Mini

FTA Classic

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Whatman FTA cards described in >400 peer-reviewed publications

Has an FTA card been used in your application? These three charts provide an illustration of how FTA cards have been used in several hundred peer-reviewed publications. From diagnostics to plant biotechnology, from human to fungal samples, the ease of sample collection and compatibility with downstream analysis techniques has endeared FTA cards to a range of disciplines. Search our online database for publications relevant to your application at: www.gelifesciences.com/dxcomponents

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How do you prepare DNA from Whatman FTA cards?

Whatman FTA

Three simple steps to pure DNA: spot, punch, and purify in less than 30 min

Spot

40 Âľl of blood produces an 8 mm spot

Punch

3 mm punch

Purity

Yields >100 ng DNA, suitable for PCR applications

Whatman FTA Elute

This procedure yields pure DNA within 30 min of punching from the card, with <5 min hands-on time. Heme protein is retained on Elute paper and does not appear in solution Multiple aliquots of eluted DNA can be used in PCR and quantitative PCR applications

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Whatman FTA cards and your whole genome amplification workflow

Genomic DNA was amplified from dried blood stored on FTA classic card after dissociation by pH denaturation (Lanes 1,3,5,7,9, and11).

STR analysis of genomic DNA amplified from FTA spotted blood

Amplified DNA was subsequently subjected to digestion with EcoR1 restriction enzyme (Lanes 2,4,6,8,10, and12). Control GenomiPhi V3 amplifications were carried out on purified blood DNA (lanes 13 and 15) and also subjected to the same restriction digests (lanes 14 and 16)

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Sample collection with Indicating Whatman FTA Elute cards prior to a HPV test

A national cervical cancer screen program in a developed economy has the following workflow 1.Woman patient has cervical smear performed by a trained professional. 2.The professionally collected cell sample is analyzed by the cytologist. a.If +ve, patient is referred to gynaecologist b.If -ve, no further need for gynaecologist c.If inconclusive or shows an indeterminate result, then a HPV test performed. d.If HPV test is +ve for any of the high-risk types, patient revisits gynaecologist and undergoes colposcopy

In a future environment, the HPV test may become the primary screen. In this case, a specific professional sample collection is not required, making it possible for the patient to collect the sample herself. Using an Indicating FTA Elute card the sample could be collected remotely and sent for lab analysis in the conventional post. If the sample is positive, the patient would visit the gynaecologist for a secondary screen. The use of FTA cards in this manner could simplify workflow, reduce healthcare costs, increase access, and expand patient base

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Simplified DNA purification workflow

Sample collection with Whatman FTA Elute eliminates the need of a DNA purification kit because DNA is eluted in solution following a simple water wash and a heat step.

This reduces the time and number of steps taken to purify DNA from your biological sample

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Safer sample handling

Consider that a biosample is not viable when it is dried(1), unlike when wet, and therefore safer than a liquid sample. Shipment of wet samples thus requires more care and attention.

Consider the security of supply when shipping biosamples refrigerated or on dry ice and the consequences of failure in the cold chain logistics workflow.

16 1 This is publicly stated by the CDC and the US Postal Service follows this guideline.


Transportation cost savings

Switching to dry sample collection can result in cost savings of up to 94%. These cost savings derive from items such as dry ice, extra packaging and documentation associated with wet sample shipments.

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Some of your questions answered

How many punches can I get from one FTA sample circle?

What size PCR amplicons can be generated with FTA Elute?

Using the 1 inch circle on the FTA Classic, Mini, Micro Cards, approximately 140 1.2 mm discs can be obtained, and over 50 of the 2.0 mm discs.

Using the standard procedure of heating punches to 95 to 98째C for 30 min. followed by vortexing, it is possible for up to 1500 bp PCR products to be amplified.

What sample types can I apply to FTA paper? A wide range including tissue cells, blood cells (human and animal), bacteria, plant cells, liver cells, buccal cells, urine, amniotic fluid, food samples, plant (leaf presses and homogenate), and tumor cells.

What techniques are FTA and FTA Elute compatible with?

~30 ng, although his will vary depending on a number of factors including % of nucleated cells in the blood and the hematocrit levels.

FTA compatibility includes PCR, whole genome amplification, and sequencing. FTA Elute compatibility includes: PCR, QPCR, and sequencing.

Yes, the cards have been approved for shipping through the U.S. Postal Service. The FTA card is designed to protect DNA, even in transport. Make sure the card is in a protective envelope to ensure safe transport.

What is the typical yield from a 3 mm punch of blood samples from FTA Elute?

Is it safe to mail FTA and FTA Elute cards with sample applied to them?

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Questions for you

How are you SHIPPING your biological samples? How are you STORING your biological samples? How are you PURIFYING DNA from your biological samples?

Did you find the content useful? Would you like to sign up for the remaining chapters in this eBook series? Visit www.gelifesciences.com/dxcomponents 19


Chapter 2

Ready-To-Goâ&#x201E;˘ and Amplification technologies

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Introduction Welcome to the second chapter of our eBook, we hope you find it useful. Parts of this chapter are extracted from the Nucleic Acid Prep Handbook (28-9624-00), which can be downloaded from www.gelifesciences.com

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We discussed in Chapter 1 that FTA cards can in many cases replace the cold chain for collection, shipping and storage of biosamples. However for PCR, most often separate PCR reagents or mixes stored in the freezer are used, and for genomic DNA preparation, column-based kits or homebrew methods are most common. In this chapter you will learn about Ready-To-Go technology and how it can be applied to simplify the amplification steps of the genomics workflow.

Important considerations during biosample amplification

Current practices include:

a)

Collection, Transport, Storage large samples

Dry/wet ice, sub-ambient temperature storage

b)

Collection, Transport, Storage small samples

Collect, ship and store on FTA cards (Chapter 1)

c)

Processing - Gene-specific

Separate PCR reagents or mixes, stored in freezer.

d)

Processing - Whole genome

Multiple genomic DNA preparations.

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Amplification of biological samples is usually done by PCR (polymerase chain reaction), and genomic DNA is usually prepared using kits or homebrew methods, but in most cases genomic DNA preparation can be replaced by whole genome amplification.

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Amplification: comparing and choosing PCR and WGA

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DNA preparation by amplification In molecular biology applications, DNA amplification can take many forms, including the ubiquitous PCR, as well as isothermal amplification using Phi29 DNA polymerase. The choice of amplification method depends on a number of factors, for example, the availability of sequence-specific primers, the intended use of the amplified product, and the quantity and quality of source material. When sequence information is available, primer- specific DNA amplification (e.g. PCR) can produce a subpopulation of DNA from the original sample. When cDNA is the objective, RNA is the starting material, so a reverse transcription reaction can be coupled with PCR (RT-PCR). Isothermal amplification (e.g. using Phi29 DNA polymerase) is an option when the amount of input material is small and inadequate sequence information is available to prepare sequence specific primers. Amplified genomic DNA and plasmid DNA can be substituted for the original unamplified DNA in most applications. For example, amplified plasmid and other circular DNA are common templates for DNA sequencing reactions. With any amplification method, quality of the output DNA correlates with the quality of the input DNA/RNA.

As is true with other molecular biology applications, DNA amplification is influenced by sample preparation. In some cases, low-quality template can be used. However, in more stringent applications such as long PCR, single-cell or low-copy-number genetic analysis, and rare allele detection, higher-quality RNA or DNA is required. Poor sample preparation can decrease the purity of DNA/RNA and may prevent accurate quantitation by spectrophotometry. Poor quality of DNA/RNA effectively decreases the concentration of usable templates, resulting in a decrease in the signal in PCR experiments and an increase in false-negative results, as well as other problems associated with incorrectly tittered template. 25


Nucleic acid amplification using PCR and RT-PCR PCR is the most commonly used method for amplifying nucleic acid samples. It has widespread utility in both scientific research and applications such as forensics and clinical work. The number of specialized PCR methods continues to grow, and a detailed discussion of all of these is outside the scope of this handbook. See references 1 and 2 for more information. The most common PCR methods are end-point PCR and real-time or quantitative PCR (qPCR). Either of these can be combined with a reverse transcriptase step (RT-PCR and RT-qPCR, respectively) to enable amplification of RNA. illustra Ready-To-Go RT-PCR Beads can be used with either total RNA or mRNA isolated using various methods. More detail on these and other experiments can be found in the Amplification Chapter of the Handbook Nucleic Acid Sample Preparation for Downstream Analyses: Principles and Methods (28-9624-00), which can be downloaded from www.gelifesciences.com

RT-PCR detection of specific mRNAs from a range of sources. Source organisms and RNAs are indicated above the lanes. The target genes for amplification and the amounts of RNA used are indicated in the table that accompanies the figure. Futher information on the different source materials can be found in the handbook (28-9624-00), Nucleic Acid Sample Preparation for Downstream Analyses: Principles and Methods (28-962400), which can be downloaded from www.gelifesciences.com

Sample preparation considerations for DNA amplification using PCR and RT-PCR The yield, fidelity, and quality of any amplification reaction can be influenced by a number of variables, one of which is sample preparation. DNA and RNA for PCR and RT-PCR, respectively, can come from a range of sources, including cell cultures, blood, bacterial cultures, and plant and animal tissues. Potential contaminants to PCR may be sample-specific or common to most sample types. Some contaminants may be introduced during sample preparation. 26


Summary of possible contaminants that may interfere with DNA amplification.

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Overview of Ready-To-Go bead technology for PCR and RT-PCR illustra Ready-To-Go PCR beads from GE Healthcare are premixed, predispensed, single-dose reactions for performing PCR amplification. Each ambient-temperature-stable bead provides the convenience of a single-dose reaction that only requires the addition of PCR primers, template, and water. Ready-To-Go PCR Beads are available in several formulations and formats, including standard PCR, “hot start” PCR, RTPCR, and RAPD analysis. All bead types use high-purity reagents, such as recombinant Taq DNA polymerase, ensuring the lowest possible levels of contaminating DNA in each bead. The bead format significantly reduces the number of pipetting steps, thereby decreasing handling errors, pipetting errors, and contamination of subsequent reactions, all of which increases overall reproducibility. Functional testing of each lot of Ready-To-Go Beads ensures that each lot will deliver similar results. PuReTaq™ Ready-To-Go PCR Beads and Ready-To-Go RT-PCR Beads are provided in 0.5 ml tubes, 0.2 ml tubes, or in a 96-well format.

How do you prepare DNA from RTG PCR beads? RTG PCR beads

Custom pack sizes and formulations of Ready-To-Go beads are also available.

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Which is the correct method for your amplification? The method used depends on many factors including: • The availability of sequence-specific primers • The intended use of the amplified product • The quantity and quality of source material. When sequence information is available, a primer-specific DNA amplification (e.g., PCR) can be performed.

When the amount of input material is small and inadequate sequence information is available to prepare sequence-specific primers, isothermal amplification using Phi29 DNA polymerase is an option. This method is also a convenient way to replenish stocks of genomic DNA.

Schematic diagram of the amplification process with the GenomiPhi amplification method. Random hexamer primers anneal to the template DNA at multiple sites. Phi29 DNA polymerase initiates replication at multiple sites on the denatured linear DNA simultaneously. As synthesis proceeds, strand displacement of complementary DNA generates new single-stranded DNA. The subsequent priming and strand displacement replication of this DNA results in the formation of double-stranded DNA.

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illustra GenomiPhi™ DNA amplification range

Phi29 DNA polymerase-based, isothermal DNA amplification is a simple, reliable alternative to other DNA amplification methods. Use amplified DNA directly for downstream applications such as sequencing, genotyping, and array CGH.

Phi29 DNA polymerase enables rapid DNA replication from multiple sites. With 3'-5' exonuclease proof-reading activity, it also results in 100-fold higher fidelity compared to Taq polymerase, for highly representative and reliable whole-genome amplification (WGA).

The one-tube, one-temperature, non-column based method offered with GenomiPhi simplifies the DNA preparation process, facilitating automation for high-throughput sample amplification.

From nanogram amounts of starting material, it can produce consistent microgram yields of high quality DNA in under an hour (illustra GenomiPhi V2). illustra GenomiPhi HY typically yields around 50 µg. GenomiPhi is now available in Ready-To-Go format.

illustra TempliPhi™ DNA amplification range Phi29-based kits for rolling-circle amplification – used for culture-free DNA preparation, to prepare DNA directly from plasmid or fosmid glycerol stocks or colonies, eliminating bacterial culture steps.

Specialized kits are available for large constructs (illustra TempliPhi Large Construct Kit), and difficult templates (illustra TempliPhi Sequence Resolver Kit).

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Isothermal DNA amplification Commercial kits for isothermal amplification are available from several suppliers. Their utility for different types of template may vary. Although isothermal amplification procedures that utilize Phi29 DNA polymerase are generally quite resistant to inhibition by mild contamination with salts, culture media, and cell debris, results may be affected by poor quality template. Therefore, DNA should be handled with care during sample preparation. If the template is a circular DNA strand, and RCA is being used, DNA nicks and strand breaks can significantly reduce amplification potential. The likelihood of nicking and strand breaks in circular templates increases as the size of the molecule increases. Because preparation of large intact circular molecules (e.g., BACs) can be quite difficult, care should be taken to avoid rapid pipetting and vortexing. If samples are linear DNA, and multiple displacement amplification is being used, the reaction relies on hybridization of multiple primers to each DNA strand for maximal amplification. Samples containing nicks and breaks can fail to amplify well, and amplification products may contain a number of rearranged products. This may have deleterious effects on downstream applications (e.g., DNA cloning or sequencing). GE Healthcare provides optimized reagents and protocols for difficult templates such as BACs (illustra TempliPhiâ&#x201E;˘ Sequence Resolver Kit) and larger templates (illustra TempliPhi Large Construct Kit).

DNA amplification using TempliPhi and GenomiPhi DNA Amplification Kits Circular DNA amplification using TempliPhi illustra TempliPhi DNA Amplification Kits from GE Healthcare are novel products developed to exponentially amplify single- or double-stranded circular DNA templates by RCA (3,4). Amplified products can be used in downstream applications such as sequencing and molecular cloning. TempliPhi DNA Amplification Kit provides RCA reagents optimized for circular DNA amplification. The reaction utilizes modified random primers to efficiently prime synthesis by the extremely processive, high-fidelity Phi29 DNA polymerase in a single-step isothermal reaction.

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DNA amplification results with illustra TempliPhi DNA Amplification Kits TempliPhi amplified DNA can be used in the same application as unamplified DNA. Plasmid, M13, and other circular DNAs are typically used as templates for sequencing. TempliPhi amplified plasmid DNA is of high quality and generates accurate sequencing results in automated cycle sequencing.

Sequence data generated from plasmid DNA amplified with illustra TempliPhi 100 Amplification Ki . Amplified DNA as subsequently sequenced using DYEnamicâ&#x201E;˘ ET Terminator Cycle Sequencing Kit and analyzed on an ABI PRISM 3100 Genetic Analyzer.

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Whole genome amplification (WGA) using GenomiPhi The GenomiPhi™ method of WGA from GE Healthcare uses Phi29 DNA polymerase, a highly processive enzyme with excellent strand-displacement activity, in combination with random sequence hexamer primers (random hexamers) to amplify DNA in an isothermal process—a thermal cycler is not required. First, random hexamers anneal to multiple sites on the denatured linear DNA template. Next, Phi29 DNA polymerase initiates replication at these sites simultaneously. As synthesis proceeds, strand displacement of upstream replicated DNA generates new single stranded DNA. Subsequent priming and strand displacement replication of this DNA produces large quantities of several-kilobase-long, double-stranded DNA suitable for genetic analyses that require high-molecular-weight product. Microgram quantities of high-molecular-weight DNA can be generated from as little as 10 ng of genomic DNA using this simple and robust procedure. DNA replication is extremely accurate due to the proofreading 3’–5’ exonuclease activity inherent in Phi29 DNA polymerase (5, 6). Note that the GenomiPhi method does not preserve epigenetic methylation patterns.

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How do you prepare DNA from RTG GenomiPhi? RTG V3 GenomiPhi kit

Summary of formats available for Ready-To-Go (RTG) amplification RTG bead format

RTG cake format

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Whatman FTA cards and your whole genome amplification workflow

Genomic DNA was amplified from dried blood stored on FTA classic card after dissociation by pH denaturation (Lanes 1,3,5,7,9, and 11). Amplified DNA was subsequently subjected to digestion with EcoR1 restriction enzyme (Lanes 2,4,6,8,10, and 12). Control GenomiPhi V3 amplifications were carried out on purified blood DNA (lanes 13 and 15) and also subjected to the same restriction digests (lanes 14 and 16).

STR analysis of genomic DNA amplified from FTA spotted blood The GenomiPhi used in these experiments is V3. This is a Ready-To-Go format, predispensed and stable at ambient temperature. GenomiPhi is also available in liquid format for added flexibility.

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Simplified DNA purification workflow

Genomic DNA replenished with GenomiPhi rather than classic genomic DNA preparation greatly reduces hands-on time, the number of steps being greatly reduced and the whole process simplified. The whole genome amplification reaction takes from 30min to around 2h depending on the protocol used.

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DNA amplification results with illustra GenomiPhi DNA Amplification Kits Amplified genomic DNA can be used in the same applications as unamplified DNA. For WGA, common applications include library construction, forensic analysis (e.g. DNA fingerprinting), genotyping (e.g. SNP analysis), PCR analysis, DNA cloning, CGH, whole genome DNA sequencing, HLA typing, and loss of heterozygosity (LOH). Microgram quantities of genomic DNA can be amplified from small amounts of input DNA using GenomiPhi V2 kit.

Amplified DNA performs similarly to unamplified genomic DNA in SNP analysis.

SNP analysis of amplified products using Affymetrix GeneChipâ&#x201E;˘ 10K human genome chip. Individual human genomic DNA (gDNA) obtained from Coriell was amplified with illustra GenomiPhi V2 kit and subjected to analysis on Affymetrix 10K SNP chip (green bar = % call rate; blue bar = % concordance).

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Do you need Custom Ready-To-Go? Stabilise and simplify GE Healthcareâ&#x20AC;&#x2122;s Ready-To-Go (RTG) technology is proven and established for stabilisation of individual enzymes/reagents and complete multiplex assays, building on nearly 20 yearsâ&#x20AC;&#x2122; development and manufacturing experience in this field. The patented technology stabilises individual proteins and reagents, as well as complete multiplex assays by providing a molecular environment that protects against conformational changes in protein structure. The result is a product that is stable at room temperature.

Benefits of RTG technology

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Ready-To-Go and GenomiPhi used in over 3000 peer-reviewed publications

These charts show how PCR, Ready-To-Go PCR and GenomiPhi WGA have been used in peerreviewed publications. Numbers represent % mention out of total number of papers. Search performed on Stanford Highwire site http://highwire.stanford.edu/cgi/ search 4 September 2013 Key words: PCR, Ready-To-Go + PCR, GenomiPhi. 40


Some of your questions answered

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Questions for you

Did you find the content useful? Would you like to sign up for the remaining chapter in this eBook series? Visit www.gelifesciences.com/dxcomponents 42


Ordering and related information

GE Healthcare’s life sciences consumables are now available from your preferred supplier. Please contact your local GE Healthcare representative for more details. Discover illustra™ nucleic acid purification; Trap™ protein sample preparation; Amersham™ blotting, labeling, and detection; Whatman™ filtration, sample prep, and diagnostics; and GE Healthcare’s new NanoVue™ Plus, the low volume spectrophotometer, for drop, measure, done simplicity. For more information visit www.gelifesciences.com

Chapter 1 Product ordering

Product literature

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Chapter 2 Product ordering

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References 1. Bartlett, J. and Stirling, D. eds. PCR Protocols (Methods in Molecular Biology). Humana Press, Inc.,New Jersey (2003). 2. Innis, M. et al. PCR Applications: Protocols for Functional Genomics. Academic Press (1999). 3. Dean, F. et al. Rapid amplification of plasmid and phage DNA using Phi 29 DNA polymerase and multiply-primed rolling circle amplification. Genome Research 11, 1095–1099 (2001). 4. Lizardi, P. et al. Mutation detection and single-molecule counting using isothermal rolling-circle amplification. Nat. Genet. 19, 225–232 (1998). 5. Estaban, J. A. et al. Fidelity of phi 29 DNA polymerase. Comparison between protein-primed initiation and DNA polymerization. J. Biol. Chem. 268, 2719–2726 (1993). 6. Nelson, J. R. et al. TempliPhi, phi29 DNA polymerase based rolling circle amplification of templates for DNA sequencing. BioTechniques 32, S44–S47 (2002).

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GE and GE Monogram are trademarks of General Electric Company. Whatman, FTA, 903, illustra, Ready-To-Go, GenomiPhi, TempliPhi, and CloneSaver are trademarks of GE Healthcare companies. GE Healthcare is the provider of the sample collection device and currently makes no claim for its suitability for a disease-specific test. The Polymerase Chain Reaction (PCR) is covered by patents owned by Roche Molecular Systems and F Hoffmann-La Roche Ltd. All third party trademarks are the property of their respective owners. FTA is a trademark of GE Healthcare companies.

All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them. GE Healthcare reserves the right, subject to any regulatory and contractual approval, if required, to make changes in specifications and features shown herein, or discontinue the product described at any time without notice or obligation. Contact your local GE Healthcare representative for the most current information. Š 2013 General Electric Company â&#x20AC;&#x201C; All rights reserved. First published March 2013 GE Healthcare UK Limited Amersham Place Little Chalfont Buckinghamshire HP7 9NA UK For local office contact information, visit www.gelifesciences.com/contact

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Sample handling and preparation within the genomics workflow