Navigation Links
Automation of TempliPhi and GenomiPhi DNA Amplification Kits on the Genesis Freedom Robotic Sample Processor

M. Adams and R. Deadman
GE Healthcare, Piscataway, NJ, US


GE Healthcare offers a range of DNA amplification products to meet all genomic DNA and sequencing template preparation needs. A combination of these kits with the Genesis Freedom™ Robotic Sample Processor (RSP) from Tecan (Fig 1) creates a total solution for high-throughput DNA preparation.

Traditional DNA purification methods often use organic solvents, are difficult to automate, or produce variable yields. The minimal number of liquid transfers in the TempliPhi™ and GenomiPhi™ DNA amplification procedures are easy to automate, with each reaction producing the same amount of high quality DNA. TempliPhi DNA Amplification Kit and TempliPhi Large Construct DNA Amplification Kit are used to prepare small (plasmids/M13) or large (fosmids/BACs) circular DNA templates, respectively, for sequencing. GenomiPhi DNA Amplification Kit prepares genomic DNA from various sources that can be directly used for genetic analyses. The flexibility of the Genesis Freedom workstation also allows easy reaction setup for downstream applications such as sequencing or PCR.


DNA amplification
TempliPhi DNA Sequencing Template Amplification Kit offers the simplest solution to plasmid DNA preparation. Microgram quantities of DNA can be prepared from picogram amounts of purified DNA, colonies, or bacterial cultures using Phi29 DNA polymerase and rolling circle amplification. This unique approach removes the need for culture growth, resulting in high quality DNA templates with less than 20 min of hands-on time.

Amplified DNA can be used directly for sequencing with any chemistry and sequencing platform. TempliPhi Large Construct Kit has been specifically optimized for amplifying large constructs such as fosmids or BACs for sequencing. Uniform quantities of DNA are amplified in all samples, eliminating the need for quantitation.

GenomiPhi DNA Amplification Kit representatively amplifies single- or double-stranded linear genomic DNA using Phi29 DNA polymerase to perform strand displacement amplification. Microgram quantities of representative DNA are produced from small amounts of starting material that can be used immediately in downstream applications.

For all kits, DNA amplification is achieved by combining the starting DNA with random hexamer primers, Phi29 DNA polymerase, dNTPs, and buffer components optimized for DNA synthesis. This reaction mixture is incubated overnight at 30 °C, during which time the available nucleotides are consumed and converted into high molecular weight fragment copies of the input DNA.


Automation
The amplification protocols are simple and allow straightforward automation. The Genesis Freedom series of RSPs from Tecan are ideal for automating DNA amplification reactions and downstream applications. The arm of the Tecan liquid handler is equipped with four or eight tips and can perform all required liquid handling steps with high sensitivity and accuracy. When the amplification reaction stops because the nucleotides in the reaction are exhausted, DNA yields are equivalent across all samples (Fig 2). Disposable tips eliminate cross-contamination concerns. With an eight-channel liquid handling arm, multiple 96-well plates of TempliPhi or GenomiPhi DNA amplification reactions can be prepared in half the time of manual pipetting. Whatever the post-amplification application (Fig 3), the open platform can be adapted. The Tecan Gemini software system allows easy protocol setup and operation of the workstation.



Conclusions
The combination of GE Healthcare’s DNA amplification products and Tecan’s Genesis Freedom workstation produces a high-performance platform for automating the labor-intensive steps traditionally associated with manual methods of DNA preparation. The simplicity of the protocols and the robotics system allow a short setup time. Amplification reactions produce consistent, high quality DNA across all samples, eliminating time-consuming quantitation and purification steps. DNA can be used directly in downstream applications with no further purification.

For more information, including protocols, please refer to the application note Automation of TempliPhi and GenomiPhi DNA Amplification Kits on the Genesis Freedom Robotic Sample Processor (63-0056-16) which is available at: www.amershambiosciences.com/promo_dm203



back to top
'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. RNAqueous-96 RNA Isolation Kit Automation Protocol
2. Scalable Automation of the TempliPhi DNA Sequencing Template Amplification and DYEnamic ET Terminator Sequencing Kits on the Sciclone ALH 3000
3. Automation of the Eppendorf Perfectprep BAC 96 DNA Kit on the Caliper Life Sciences Sciclone ALH 3000
4. Oragene and Whole Genome Amplification with GenomiPhi
5. GenomiPhi: protocol for BAC amplification and fabrication of slides for array CGH
6. GenomiPhi amplification of FFPE tissue DNA for array CGH
7. prostar RT-PCR Systems for Robust High-Fidelity RNA Amplification
8. Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
9. Greater Amplification Specificity with New Hot Start PCR Enzyme
10. Enhanced Amplification of Long Targets with PfuTurbo DNA Polymerase
11. Optimizing pfuturbo DNA Polymerase Amplification Reactions with Perfect Match PCR Enhancer
Post Your Comments:
(Date:8/28/2014)... Sterlitech is proud to announce that it ... stable of products . These additional products increase ... membrane filters with surface charges. , “We ... our membrane process testing equipment, and correspondingly, a wider ... explains Sterlitech President Mark Spatz. “The addition of ...
(Date:8/28/2014)... (PRWEB) August 27, 2014 Scientists, researchers, ... for SPIE Laser Damage 2014 symposium. Marking ... for high-power lasers will run 14-17 September. The event ... optics and photonics . , The premier conference for ... laser damage to optical materials will engage researchers and ...
(Date:8/28/2014)... PrimeSource Building Products, Inc., a ... SoundConnect to support their communication needs with ... web conferencing platform GlobalMeet powered by SoundConnect. The ... SoundConnect delivers award winning “results driven” business ... will be utilizing GlobalMeet powered by SoundConnect ...
(Date:8/27/2014)... August 27, 2014 Varian Medical Systems ... its current Salt Lake City manufacturing facility and allow ... Utah to ultimately create 1,000 new full time jobs ... , “Utah is home to hardworking people with immense ... credited with Varian’s success in the manufacturing of our ...
Breaking Biology Technology:Sterlitech Corporation Enhances Its Selection of Flat Sheet Membrane Filters 2Laser Damage to Draw Researchers, Engineers, Scientists to 46th Annual Conference 2Laser Damage to Draw Researchers, Engineers, Scientists to 46th Annual Conference 3PrimeSource Chooses SoundConnect as Collaboration Provider 2Varian Medical Systems Groundbreaking on Facility Expansion Triggers Influx of New Jobs to Salt Lake 2Varian Medical Systems Groundbreaking on Facility Expansion Triggers Influx of New Jobs to Salt Lake 3
... MISSISSAUGA, ON, Dec. 14 /PRNewswire-FirstCall/ - YM BioSciences Inc. ... life sciences product development company that identifies and advances ... stages of development, today announced that it has priced ... a price to the public of $1.60 per share. ...
... CAMBRIDGE, Mass., Dec. 14, 2010 Tolerx, Inc., a ... and cancer by modulating T cell activity, today announced ... to assess the safety, tolerability, pharmacokinetic, and pharmacodynamic profiles ... immune system by enabling T cells to more effectively ...
... Cambridge, Mass., December 6, 2010 Utilizing a century-old ... at Harvard University have demonstrated, for the first time, ... collaboration with researchers from Hamamatsu Photonics in Hamamatsu City, ... University of Magdeburg, Germany, the advance has a wide ...
Cached Biology Technology:YM BIOSCIENCES PRICES $40 MILLION PUBLIC OFFERING 2YM BIOSCIENCES PRICES $40 MILLION PUBLIC OFFERING 3Tolerx Initiates Phase 1 Clinical Trial of TRX518, a First-in-Class anti-GITR Antibody for the Treatment of Cancer 2Tolerx Initiates Phase 1 Clinical Trial of TRX518, a First-in-Class anti-GITR Antibody for the Treatment of Cancer 3Capasso lab demonstrates highly unidirectional 'whispering gallery' microlasers 2
(Date:8/28/2014)... to Research Careers) Program has announced the travel award ... October 18 22, 2014 in San Diego, California. ... of students, post doctorates and scientists from underrepresented groups ... to encourage the participation of young scientists at the ... conferred 16 awards totaling $29,600., The FASEB MARC Program ...
(Date:8/28/2014)... Researchers at the University of Minnesota have developed ... to be used for muscle regeneration research as well ... FSHD. , The research is published in the current ... There is no treatment for FSHD, which is thought ... muscular dystrophy. FSHD is an unusual genetic disorder because, ...
(Date:8/28/2014)... support of peer groups and clinicians is critical ... to recent University of Georgia research. , A ... County area determined that role models for successful ... of infants. , "Mothers who received that support ... said study co-author Alex Anderson, an associate professor ...
Breaking Biology News(10 mins):UMN researchers find animal model for understudied type of muscular dystrophy 2Breastfeeding study shows need for effective peer counseling programs 2
... low intake of folate and vitamin B12 increases the ... among nearly 3,000 middle-aged and elderly Finnish subjects. On ... an increased risk for the metabolic syndrome. Based on ... separate depressive subtypes with different etiologies in terms of ...
... This press release is available in Spanish . ... of amino acids and they play a fundamental role in ... in genes; they, therefore, carry out numerous functions in an ... material), bioregulating (they form part of enzymes) and a long ...
... how life began on a molecular level has been a ... on a possible mechanism by which life may have gotten ... early Earth. Researchers have proposed several competing theories for ... before the first genes or living cells came to be. ...
Cached Biology News:Lack of nutrients and metabolic syndrome linked to different subtypes of depression 2Using computational biology for the annotation of proteins 2Using computational biology for the annotation of proteins 3Model sheds light on the chemistry that sparked the origin of life 2
... Tissue Staining Kits are intended for localization ... histological and cytological specimens. These kits may ... by either R&D Systems or by other ... formation of the Avidin-Biotin Complex (ABC) with ...
... Designed specifically for high efficiency ... Low toxicity reagent proven to ... range of cell types. Results ... 95% of target gene expression. ...
... Purification Protocols covers a wide range of ... and experienced researchers in all areas of ... from the books preface illustrates just how ... research library., The chapters introduce the theoretical ...
... jetSI-ENDO is an easy-to-use siRNA transfection reagent ... endogenous genes in a variety of cell ... with an optimized, straightforward protocol. High levels ... obtained following transfection of 20 to 100 ...
Biology Products: